Compounds for treating tumors

ABSTRACT

The invention provides compounds of formula (I): 
                         
wherein E, A, B′, R 6 , R 7 , R 8 , and R 9  are defined in the specification which compounds exhibit anticancer activity and are useful for treating cancer.

This application claims priority from copending provisional applicationSer. No. 60/493,841, filed Aug. 8, 2003, the entire disclosure of whichis hereby incorporated by reference.

FIELD OF THE INVENTION

The present invention relates to new hemiasterlin derivatives and saltsthereof which demonstrate anti-mitotic and cytotoxic activity and areuseful for treating cancer. The present invention further relates to amethod of treating or inhibiting the growth of cancerous tumour cellsand associated diseases in a mammal and additionally relates topharmaceutical compositions containing compounds of the invention andsalts thereof, and novel processes for preparing compounds of theinvention and salts thereof.

BACKGROUND OF THE INVENTION

Drug resistance is a major impediment in cancer chemotherapy. Patientsmay acquire resistance during multiple cycles of therapy. Alternatively,patients may not respond at the onset of therapy (inherent resistance).Resistance to virtually all approved cancer cytotoxic chemotherapy hasbeen reported including antimitic agents (paclitaxel and docetaxel,vinblastine, vincristine or vinorelbine), topoisomerase inhibitors(etoposide, teniposide, topotecan, camptothecin, doxorubicin andduanorubicin), antimetabolities (methotrexate, 5-flurouracil,gemcitabine), alkylating agents (melphalan, chlorambucil), and other DNAdamaging agents (cisplatin and its analogs). (Goldstein, L. J., Ozols,R. F. Anticancer Drug Resistance, p. 294. Boston: Kluwer AcademicPublishers, 1994.) The mechanisms mediating resistance are highlydiverse. Finding and developing chemical compounds having novelstructures for use in tumor therapy and which overcome resistance isimportant.

Hemiasterlins are natural products derived from sponges that inducemicrotubule depolymerization, G₂/M cell cycle arrest, and ultimatelycell death.

Finding and developing chemical compounds having novel structures foruse in tumor therapy and which overcome resistance is important.(Anderson, H. J., Coleman, J. E., Andersen, R. J., Roberge, M. Cytotoxicpeptides hemiasterlin, hemiasterlin A and hemiasterlin B induce mitoticarrest and abnormal spindle formation, Cancer Chemother. Pharmacol. 39:223-226, 1997. Talpir, R., Benayahu, Y., Kashman, Y., Pannell, L.,Schleyer, M. Hemiasterlin and geodiamolide TA: two new cytotoxicpeptides from the marine sponge hemiasterellaminor (kirkpatrick),Tetrahedron Letters 35: 4453-4456, 1994). Hemiasterlins in cancertherapy have been reported (WO 99/32509, WO 96/33211 and U.S. Pat. No.6,153,590). Further, Hemiasterlin may be obtained from marine sponges(U.S. Pat. No. 5,661,175 (1997) and U.S. Pat. No. 6,153,590 (2000)) orsynthesized (Andersen, R. J., Coleman, J. E. Tetrahedron Letters 38:317-320, 1997). Synthetic analogs of hemiasterlin have also beenprepared (WO 99/32509), and Hemiasterlin and the synthetic analogsthereof are reported to have cytotoxic and anti-mitotic activities.

The mechanisms that may mediate resistance to known antimitotic agentsinclude drug efflux pumps (MDR1 and possibly MXR), tubulin mutations,alternative isotypes expression of tubulin isomers, alteration in theexpression or function of genes that mediate apoptosis (e.g. p53 andbcl-2), and overexpression of growth factors such as HER-2. (Rowinsky,E. K., Tolcher, A. W. Antimicrotubule agents. In: J. Devita, V. T.,Hellman, S., Rosenberg, S. A. (ed.) Cancer Principles and Practice, 6thedition edition, pp. 431-452. Philadelphia: Lippincott Williams andWilkins, 2001) Resistance mediated by the multidrug drug resistancegene, MDR-1, has been intensively studied, mainly because it isfrequently encountered in experimental models. (Greenberger, L. M.,Cohen, D., and Horwitz, S. B. In vitro models of multiple drugresistance. In: a. R. F. O. L. J. Goldstein (ed.) Anticancer DrugResistance, pp. 69-106. Norwell, Mass.: Kluwer Academic Publishers,1994) MDR-1 is implicated in resistance to anti-microtubule agentssince: 1) selection of tissue culture cells for resistance to vincaalkaloids or taxanes leads to marked overexpression of MDR-1, 2) cellsthat overexpress MDR-1 have low drug accumulation of taxanes or vincaalkaloids, 3) transfection of cells with MDR-1 induces resistance tothese agents, 4) photoaffinity probes for vinca alkaloids or taxanesbind to the MDR-1 gene product, P-glycoprotein, 5) transgenic micedevoid of MDR gene family members have altered pharmacokinetic profilesfor taxanes and 5) agents that inhibit P-glycoprotein resensitizeresistant cells to taxanes or vinca alkaloids. The clinical relevance ofMDR-1 overexpression is not clear in most solid tumor types and itsassociation with lack of patient response or poor prognosis iscontroversial. (Bradshaw, D. M., Arceci, R. J. Clinical relevance oftransmembrane drug efflux as a mechanism of multidrug resistance, J.Clin. Oncol. 16: 3674-3690, 1998) Nevertheless, overexpression of MDR1has been clearly associated with response to chemotherapy and prognosisin leukemias. Low level resistance to vinca alkaloids (but not taxanes)has also been found in cells transfected with another efflux pump, MRP.(Breuninger, L. M., Paul, S., Gaughan, K., Miki, T., Chan, A., Aaronson,S. A., Kruh, G. D. Expression of multidrug resistance-associated proteinin NIH/3T3 cells confers multidrug resistance associated with increaseddrug efflux and altered intracellular drug distribution., Cancer Res.55: 5342-5347, 1995; Zaman, G. J. R., Flens, M. J., van Leusden, M. R.,de Haas, M., Mulder, H. S., Lankelma, J., Pinedo, H. M., Scheper, R. J.,Baas, F., Broxterman, H. J., and Borst, P. The human multidrugresistance-associated protein MRP is a plasma membrane drug-efflux pump,Proc. Natl. Acad. Sci. USA. 91: 8822-8826,1994.) Tubulin mutations havebeen found in cells selected for resistance to agents that polymerizemicrotubules, paclitaxel or epothilones. (Giannakakou, P., Gussio, R.,Nogales, E., Downing, K. H., Zaharevitz, D., Bolbuck, B., Poy, G.,Sackett, D., Nicolauo, K. C., Fojo, T. A common pharmacophore forepithilone and taxanes: molecular basis for drug resistance conferred bytubulin mutations in human cancer cells, Proc. Natl. Acad. Sci. USA. 97:2904-2909, 2000; Giannakakou, P., Sackett, D. L., Kang, Y.-K., Zhan, A.,Buters, J. T., M., Fojo, T., Poruchynsky, M. S. Paclitaxel-resistanthuman ovarian cancer cells have mutant b-tubulins that exhibit impairedpaclitaxel-driven polymerization, J. Biol. Chem. 272: 17118-17125,1997.) For paclitaxel resistance of this type, selection must be donewith paclitaxel in the presence of an MDR-1 inhibitor to avoid thepreferential overexpression of MDR1. Based on crystallographic data andmolecular modeling of tubulin, the mutations occur in regions of tubulinthought to interact with taxanes. (Giannakakou, P., Gussio, R., Nogales,E., Downing, K. H., Zaharevitz, D., Bolbuck, B., Poy, G., Sackett, D.,Nicolauo, K. C., Fojo, T. A common pharmacophore for epithilone andtaxanes: molecular basis for drug resistance conferred by tubulinmutations in human cancer cells, Proc. Natl. Acad. Sci. USA. 97:2904-2909, 2000.) While clinical significance is still being evaluated,one report found that 33% of patients with non-small cell carcinomas hadtumors with tubulin mutations and such mutations are correlated withpoor response to paclitaxel therapy. (Monzo, M., Rosell, R., Sanchez, J.J., Lee, J. S., O'Brate, A., Gonzalez-Larriba, J. L., Alberola, V.,Lorenzo, J. C., Nunez, L., Ro, J. Y., Martin, C. Paclitaxel resistancein son-small cell lung cancer associated with beta-tubulin genemutations, J. Clin. Oncol. 17: 1786-179, 1999.) Differential expressionof tubulin isoforms has been found in some cell lines selected forpaclitaxel or vinca alkaloid resistance. (Burkart, C. A., Kavallaris,M., Horwitz, S. B. The role of b-tubulin isotypes in resistance toantimitotic drugs, Biochim. Biophys. Acta. 1471: 01-09, 2001.) Theclinical association with isotype alterations has not been fullystudied, but alterations in isotype expression in patients resistant topaclitaxel have been found. (Kavallaris, M., Kuo, D. Y-.S., Burkhart, C.A., Regf, D. L., Norris, M. D., Haber, M., Horwitz, S. B.Taxol-resistant epithelial ovarian tumors are associated with alteredexpression of specific b-tubulin isotypes, J. Clin. Invest. 100:1282-1293,1997.)

There is still a need for novel compounds which have chemical structuresother than paclitaxel, which further have anticancer activity and inparticular which have antimicrotubule activity against resistant celllines and are useful for the treatment of cancer.

BRIEF SUMMARY OF THE INVENTION

In accordance with the present invention, there is provided compoundsrepresented by Formula I:

wherein:

A is selected from the group consisting of an alkyl moiety of 1 to 10carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and a cyclichydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atoms mayoptionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygen atoms,and 0 to 4 sulfur atoms, and the carbon atoms are optionally substitutedwith: ═O, ═S, OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂, —NR₁₀H,—N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —CI, —F, —CN, —CO₂H, —CHO,—COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H,—SOR₁₀, —SO₂R₁₀, wherein R₁₀ is a alkyl moiety of 1 to 10 carbon atoms,alkenyl moiety of 2 to 10 carbon atoms, aryl and a cyclic hydrocarbonmoiety of 3 to 10 carbon atoms, aryl-R— and heteroaryl-R; or

A is OR, S(O)R, S(O)₂R, SO₂NR₂, NR₁R₂ or N₃;

R is selected from the group consisting of H, an alkyl moiety of 1 to 18carbon atoms, alkenyl moiety of 2 to 18 carbon atoms, aryl and a cyclichydrocarbon moiety of 3 to 18 carbon atoms, wherein carbon atoms mayoptionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygen atoms,and 0 to 4 sulfur atoms, and the carbon atoms are optionally substitutedwith: ═O, ═S, OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂, —NR₁₀H,—N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —CI, —F, —CN, —CO₂H, —CO₂R₁₀,—CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H,—SOR₁₀, or —SO₂R₁₀;

B′ is O or H₂;

E is the moiety

-   -   or aryl, a 5 to 14-membered monocyclic, bicyclic or tricyclic        saturated or unsaturated hydrocarbon ring moiety wherein carbon        atoms may optionally be replaced with 0 to 4 nitrogen atoms, 0        to 4 oxygen atoms, and 0 to 4 sulfur atoms wherein the carbon        atoms may optionally be substituted with: R, ═O, ═S, OH, —OR₁₀,        —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀,        —NR₁₀COR₁₀, —I, Br, —Cl, —F, —CN, —CO₂H, —CHO, —COR₁₀, —CONH₂,        —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, or        —SO₂R₁₀;    -   R₁ is selected from the group consisting of H, an alkyl moiety        of 1 to 10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms,        aryl and a cyclic hydrocarbon moiety of 3 to 10 carbon atoms,        wherein carbon atoms may optionally be replaced with 0 to 4        nitrogen atoms, 0 to 4 oxygen atoms, and 0 to 4 sulfur atoms,        and the carbon atoms may optionally be substituted with: ═O, ═S,        OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂,        —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —Cl, —F, —CN, —CO₂H, —CHO, —COR₁₀,        —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H,        —SOR₁₀, or —SO₂R₁₀;

R₂ is selected from the group consisting of H, an alkyl moiety of 1 to10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and acyclic hydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atomsmay optionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygenatoms, and 0 to 4 sulfur atoms, and the carbon atoms may optionally besubstituted with: ═O, ═S, OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂,—NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —Cl, —F, —CN, —CO₂H,—CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H,—SOR₁₀, or —SO₂R₁₀;

R₁ and R₂ together may optionally form a ring of 3 to 7 carbon atomswherein carbon atoms may optionally be replaced with 0 to 2 nitrogenatoms, 0 to 2 oxygen atoms and 0 to 2 sulfur atoms;

R₃ is selected from the group consisting of H, an alkyl moiety of 1 to10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and acyclic hydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atomsmay optionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygenatoms, and 0 to 4 sulfur atoms, and the carbon atoms may optionally besubstituted with: ═O, ═S, OH, —OR₁₀, —O₂CR₁₀, —SH, —SOCR₁₀, —NH₂,—NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —CI, —F, —CN, —CO₂H,—CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀,—NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀;

R₄ is selected from the group consisting of H, an alkyl moiety of 1 to10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and acyclic hydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atomsmay optionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygenatoms, and 0 to 4 sulfur atoms, and the carbon atoms may optionally besubstituted with: ═O, ═S, OH, —OR₁₀, —O₂CR₁₀, —SH, —SOCR₁₀, —NH₂,—NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —CI, —F, —CN, —CO₂H,—CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀,—NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀;

R₃ and R₄ together may optionally form a ring of 3 to 7 carbon atomswherein carbon atoms may optionally be replaced with 0 to 2 nitrogenatoms, 0 to 2 oxygen atoms and 0 to 2 sulfur atoms;

R₅ is selected from the group consisting of H, OH, NHR, SH, aryl,heteroaryl, an alkyl moiety of 1 to 10 carbon atoms, alkenyl moiety of 2to 10 carbon atoms and a cyclic hydrocarbon moiety of 3 to 10 carbonatoms, wherein carbon atoms may optionally be replaced with 0 to 4nitrogen atoms, 0 to 4 oxygen atoms, and 0 to 4 sulfur atoms, and thecarbon atoms may optionally be substituted with: ═O, ═S, OH, —OR₁₀,—O₂CR₁₀, —SH, —SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I,Br, —Cl, —F, —CN, —CO₂H, —CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀,—CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀;

R₅ and A may optionally form a ring of 5 to 7 carbon atoms whereincarbon atoms may optionally be replaced with 0 to 2 nitrogen atoms, 0 to2 oxygen atoms, and 0 to 2 sulfur atoms;

R₆ is selected from the group consisting of H, an alkyl moiety of 1 to10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and acyclic hydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atomsmay optionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygenatoms, and 0 to 4 sulfur atoms, and the carbon atoms may optionally besubstituted with: ═O, ═S, OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂,—NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —Cl, —F, —CN, —CO₂H,—CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀,—NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀;

R₇ is selected from the group consisting of H, an alkyl moiety of 1 to10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and acyclic hydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atomsmay optionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygenatoms, and 0 to 4 sulfur atoms, and the carbon atoms may optionally besubstituted with: ═O, ═S, OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂,—NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —Cl, —F, —CN, —CO₂H,—CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀,—NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀;

R₈ is selected from the group consisting of H, an alkyl moiety of 1 to10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and acyclic hydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atomsmay optionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygenatoms, and 0 to 4 sulfur atoms, and the carbon atoms may optionally besubstituted with: ═O, ═S, OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂,—NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —Cl, —F, —CN, —CO₂H,—CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀,—NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀;

R₉ is selected from the group

W′ is selected from the group consisting of SO₂R₁₆, SO₃R₁₄, SO₂NR₁₄R₁₅,P(O)(OR₁₄)(OR₁₅), CN, OH, tetrazole, a moiety

and SO₂NRR where the R groups may form a 4 to 8 membered ring whereincarbon atoms may optionally be replaced with 0 to 2 nitrogen atoms, 0 to2 oxygen atoms and 0 to 2 sulfur atoms;

R₁₄ and R₁₅ are independently selected from the group H and alkyl of 1to 6 carbon atoms;

R₁₆ is alkyl of 1 to 6 carbon atoms;

D is O or OH;

Z and Y may optionally form a ring of 5 to 7 carbon atoms wherein carbonatoms may optionally be replaced with 0 to 2 nitrogen atoms, 0 to 2oxygen atoms, and 0 to 2 sulfur atoms;

Y is an alkyl moiety of 1 to 10 carbon atoms optionally substituted withR, ArylR-, or X, or an alkenyl moiety of 2 to 10 carbon atoms optionallysubstituted with R, ArylR-, or X;

Z is defined as a moiety selected from the group consisting of: H, alkylof 1 to 6 carbon atoms, —NRN(R)₂, R, aryl, heteroaryl, aralkyl, —OR,—SH, —SR, —NH₂, —NHR, —NROR, —N(R)₂, NH—NH₂, NRR where the R groups mayform a 4 to 8 membered ring wherein carbon atoms may optionally bereplaced with 0 to 2 nitrogen atoms, 0 to 2 oxygen atoms, and 0 to 2sulfur atoms, —NHCH(R₁₁)COOH; and —NRCH(R₁₁)COOH, wherein R₁₁ is amoiety having the formula: R, or —(CH₂)_(n)NR₁₂R₁₃, wherein n=1-4 andR₁₂ and R₁₃ are independently selected from the group consisting of: H;R; and —C(NH)(NH₂); or Z is

selected from moieties of the formula

the dotted line is an optional bond;

G₁ is selected from O, N and S;

m is an integer of 1 to 3;

u is an integer of 0-5;

R₁₇ is phenyl, or O—(CH₂)_(n)phenyl;

R₁₈ is H or OH;

R₁₉ is selected from a bond, an alkyl moiety of 1 to 10 carbon atomsoptionally substituted with an alkyl moiety of 1 to 10 carbon atoms, andalkoxy of 1 to 10 carbon atoms;

R₂₀ is selected from OR₁₄, NH—R₂₁, a moiety of the formula

and a moiety of the formula

R₂₁ is an alkyl moiety of 1 to 10 carbon atoms optionally substitutedwith aryl and heteroaryl;

R₂₂ is an alkyl moiety of 1 to 10 carbon atoms optionally substitutedwith aryl;

-   -   X is defined as a moiety selected from the group consisting of:        —OH, —OR, ═O, ═S, —O₂CR, —SH, —SR, —SOCR, —NH₂, —NHR, —N(R)₂,        —NHCOR, —NRCOR, —I, —Br, —Cl, —F, —CN, —CO₂H, —CO₂R, —CHO, —COR,        —CONH₂, —CONHR, —CON(R)₂, —COSH, —COSR, —NO₂, —SO₃H, —SOR, and        —SO₂R;

Aryl is defined as an aromatic hydrocarbon moiety having 6, 10 or 14carbon atoms, optionally substituted with R or Z;

-   -   Heteroaryl means a 5- or 6-membered heterocyclic ring, which may        be fused to another 5- or 6-membered heterocyclic ring,        especially heteroaromatic rings which contain 1 to 3 heteroatoms        independently selected from O, N and S optionally substituted        with R or X or fused to a cyclic hydrocarbon moiety of 3 to 10        carbon atoms;

provided when A is NR₁R₂, B′ is O, D is OH and R₁, R₂, R₃, R₄, R₅, R₆,R₇and R₈ are independently selected from the group consisting of H, analkyl moiety of 1 to 10 carbon atoms, alkenyl moiety of 2 to 10 carbonatoms, aryl and a cyclic hydrocarbon moiety of 3 to 10 carbon atoms, 0to 4 nitrogen atoms, 0 to 4 oxygen atoms, and 0 to 4 sulfur atoms, andthe carbon atoms may optionally be substituted with: ═O, ═S, OH, —OR₁₀,—O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀,—NR₁₀COR₁₀, —I, Br, —Cl, —F, —CN, —CO₂H, —CO₂R₁₀, —CHO, —COR₁₀, —CONH₂,—CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀;

then Z is not a moiety selected from the group consisting of: —OH, —OR;—SH; —SR; —NH₂; —NHR; —N(R)₂; —NHCH(R₁₁)COOH; and —NRCH(R₁₁)COOH,wherein R₁₁ is a moiety having the formula: R, or —(CH₂)_(n)NR₁₂R₁₃,wherein n=1-4 and R₁₂ and R₁₃ are independently selected from the groupconsisting of: H; R; and —C(NH) (NH₂);

or pharmaceutically acceptable salts thereof.

Accordingly, an object of this invention is to provide compounds whichovercome the above described limitations in cancer treatment byproviding a method for treating tumors that are resistant to currentlymarketed antimitotic agents. In particular, the method is useful intumor cells that overexpress MDR-1, MXR, or MRP.

Further, an additional object of this invention provides a method fortreating or inhibiting multiple drug resistant tumors in a mammal inneed thereof. In particular, this application demonstrates thatcompounds of the invention are effective in tumor cells that expressmultiple drug resistance and that have inherent or acquired resistancein drug resistant tumors.

The present invention also provides a pharmaceutical composition whichcomprises a compound of this invention in combination or associationwith one or more pharmaceutically acceptable carriers. In particular,the present invention provides a pharmaceutical composition whichcomprises an effective amount of a compound of this invention and andone or more pharmaceutically acceptable carriers.

Definitions:

The term alkyl means a saturated linear, or branched, hydrocarbon moietyof 1 to 10 carbon atoms. In some embodiments of the invention the moietymay optionally be 1 to 18 carbon atoms or 1 to 6 carbon atoms. Examplesinclude methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, sec-butyl,tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl, n-hexyl,2-methylphenyl, 2,2-dimethylbutyl, n-heptyl, 2-methylhexyl, and the likeunless otherwise specified. The carbon atoms may optionally be replacedwith 0 to 4 nitrogen atoms, 0 to 4 oxygen atoms, and 0 to 4 sulfuratoms, and the carbon atoms are optionally substituted with: ═O, ═S, OH,—OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀,—NR₁₀COR₁₀, —I, Br, —Cl, —F, —CN, —CO₂H, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀,—CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, —SO₂R₁₀, wherein R₁₀ isa an alkyl moiety of 1 to 10 carbon atoms, alkenyl moiety of 2 to 10carbon atoms, aryl and a cyclic hydrocarbon moiety of 3 to 10 carbonatoms, aryl-R— and heteroaryl-R.

The term alkenyl means an unsaturated linear, or branched, hydrocarbonmoiety means a moiety of 2 to 10 carbon atoms containing at least onecarbon-carbon double bond, each double bond being independently cis,trans or a nongeometric isomer wherein carbon atoms may optionally bereplaced with 0 to 4 nitrogen atoms, 0 to 4 oxygen atoms, and 0 to 4sulfur atoms, and the carbon atoms are optionally substituted with: ═O,═S, OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂,—NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —Cl, —F, —CN, —CO₂H, —CHO, —COR₁₀, —CONH₂,—CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, —SO₂R₁₀,wherein R₁₀ is an alkyl moiety of 1 to 10 carbon atoms, alkenyl moietyof 2 to 10 carbon atoms, aryl and a cyclic hydrocarbon moiety of 3 to 10carbon atoms, aryl-R— and heteroaryl-R.

The term cyclic hydrocarbon moiety means a saturated or unsaturatedcyclic hydrocarbon moiety of 3 to 10 carbon atoms, a monocycliccycloalkyl or cycloalkenyl ring of 3 to 10 carbon atoms wherein carbonatoms may optionally be replaced with 0 to 4 nitrogen atoms, 0 to 4oxygen atoms, and 0 to 4 sulfur atoms, and the carbon atoms areoptionally substituted with: ═O, ═S, OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀,—SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —Cl, —F,—CN, —CO₂H, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀,—NO₂, —SO₃H, —SOR₁₀, —SO₂R₁₀, wherein R₁₀ is an alkyl moiety of 1 to 10carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and a cyclichydrocarbon moiety of 3 to 10 carbon atoms, aryl-R— and heteroaryl-R. Insome embodiments of the invention the cyclic hydrocarbon may optionallybe a 5 to 14-membered monocyclic, bicyclic or tricyclic saturated orunsaturated hydrocarbon ring moiety wherein carbon atoms may optionallybe replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygen atoms, and 0 to 4sulfur atoms wherein the carbon atoms may optionally be substitutedwith: R, ═O, ═S, OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂, —NR₁₀H,—N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —Cl, —F, —CN, —CO₂H, —CHO,—COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H,—SOR₁₀, or —SO₂R₁₀;

Aryl is defined as an aromatic hydrocarbon moiety having 6, 10 or 14carbon atoms, preferably 6 to 10 carbon atoms optionally substitutedwith R or X. In particular an aromatic moiety is selected from the groupconsisting of: phenyl, naphthyl, anthracyl, and phenanthryl optionallysubstituted with R or X;

The term heteroaryl means a 5- or 6-membered heterocyclic ring, whichmay be fused to another 5- or 6-membered heterocyclic ring, especiallyheteroaromatic rings which contain 1 to 3 heteroatoms selected from O, Nand S optionally substituted with R or X or fused to a cyclichydrocarbon moiety of 3 to 10 carbon atoms. Exemplary heteroaromaticrings include but are not limited to thienyl, furyl, indolyl, pyrrolyl,thiophenyl, benzofuryl, benzothiophenyl, quinolyl, isoquinolyl,imidazolyl, thiazolyl, oxazolyl, and pyridyl.

The term alkoxy means an alkyl-O— group in which the alkyl group is ashereinbefore defined. Exemplary alkoxy groups include but are notlimited to methoxy, ethoxy, n-propoxy, 1-propoxy, n-butoxy and t-butoxy.

Aralkyl as used herein means an aryl-alkyl group in which the aryl andalkyl group are previously defined. Exemplary non-limiting examples ofaralkyl groups include benzyl and phenethyl.

The term phenyl as used herein refers to a 6-membered carbon aromaticring.

Preferably the recitation of a compound of Formula (I) herein covers allpossible salts of the compound, and denotes all possible isomerspossible within the structural formula given for such compound,including geometrical and optical isomers. Unless otherwise stated,materials described herein comprising a compound for which isomersexist, are to be regarded as covering individual isomers, and, mixturesof isomers including racemic mixtures.

Embodiments of this invention include the following:

in a compound of Formula (I), the following substituents alone, or incombination, are preferred:

(a) R₁ is H, methyl, ethyl, propyl, or n-butyl and R₂ is methyl, ethyl,propyl, or

 n-butyl; or, where R₁ and R₂ taken together with the nitrogen atom towhich they are attached form a 3 to 8 membered ring; more preferably R₁is H and R₂ is CH₃;

(b) preferably no more than one of R₃ and R₄ is H; more preferably, R₃and R₄ are independently: methyl, ethyl, n-propyl or n-butyl, or, whereR₃ and R₄ are joined together to form a β-cyclopropyl, β-cyclobutyl,β-cyclopentyl or β-cyclohexyl ring; most preferably R₃ and R₄ are eachmethyl;

(c) R₅: is cyclohexyl; Aryl in the definition of R₅ is preferablyphenyl, or naphthyl; heteroaryl in the definition of R₅ is preferablythienyl, or indolyl; preferably R₅ is phenyl, or indolyl; mostpreferably R₅ is phenyl;

(d) preferably E is phenyl, adamantly, or1,2,3,4-tetrahydro-1-naphthalenyl;

(e) R₆ and R₈ are independently: H or methyl, more preferably R₆ is Hand R₈ is methyl;

(f) R₇ is a three to six carbon, branched alkyl moiety; more preferablyR₇ is —C(CH₃)₃;

(g) in R₉, Z is a moiety

where m is 1 or 2; R₁₉ is a bond or a saturated alkyl chain of 3 or 4carbon atoms optionally substituted with alkoxy of 1 to 3 carbon atoms;R₂₀ is a moiety

where R₂₂ is alkyl of 1 to 3 carbon atoms substituted with aryl andthiazole.

(h) R₁₉ is an alkyl moiety of 1 to 6 carbon atoms optionally substitutedwith a alkyl moiety of 1 to 6 carbon atoms, and alkoxy of 1 to 6 carbonatoms straight or branched chain alkyl;

(i) in R₉ where Z is preferably, morpholino, piperazinyl, imidazole,phenyl, N-methyl-piperazinyl, N-benzyl-piperazinyl or pyrrolidin-1-ylcarboxylic acid or alkyl ester;

(j) R₉ is preferably —C(R₁₅)—C═C(R₁₆)C(O)-Z wherein R₁₅ is methyl,ethyl, n-propyl, isopropyl, tert-butyl, iso-butyl, or sec-butyl and R₁₆.is H, methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl orsec-butyl;

(k) R₉ is preferably

R₉ is more preferably;

R₉ is most preferably;

Additionally preferred compounds of the invention are those of Formula(I) wherein:

A is NR₁R₂;

R₁ is methyl;

R₂ is H;

R₃ and R₄ are methyl;

R₅ is selected from the group consisting of H, an alkyl moiety of 1 to10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and acyclic hydrocarbon moiety of 3 to 10 carbon atoms, said carbon atomsbeing optionally substituted with: ═O, ═S, OH, —OR₁₀, —O₂CR₁₀, —SH,—SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —Cl, —F,—CN, —CO₂H, —CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH,—COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀, wherein R₁₀ is an alkyl moietyof 1 to 10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryland a cyclic hydrocarbon moiety of 3 to 10 carbon atoms, aryl-R— andheteroaryl-R;

R₇ is preferably t-butyl;

R₉ is preferably —C(R₁₅)—C═C(R₁₆)C(O)-Z wherein R₁₅ is methyl, ethyl,n-propyl, isopropyl, tert-butyl, iso-butyl, or sec-butyl and R₁₆ is H,methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl or sec-butyl;

R₉ is:

R₉ is more preferably

R₉ is most preferably

The most preferred absolute configurations of compounds of Formula (I)wherein the absolute configurations of moieties a, b and c of Formula(I)

are selected from:

a b c S S S R S S and S S R.

Additionally preferred compounds of the invention are those of Formula(I) and pharmaceutically acceptable salts thereof wherein:

W′ is selected from the group consisting of tetrazole, SO₃C₂H₅, SO₃H,P(O)(OCH₃)₂; P(O)(OH)(OCH₃), P(O)(OH)₂, P(O)(OC₂H₅)₂,

Specifically preferred compounds of the invention includingpharmaceutically acceptable salts thereof include:

-   N,β,β,3-tetramethyl-L-phenylalanyl-N¹—[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4    -yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,β,β,3,4-Pentamethyl-L-phenylalanyl-N¹—[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-    4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,β,β,3,5-Pentamethyl-L-phenylalanyl-N¹—[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N′,3-dimethyl-L-valinamide,-   N,β,β,3,4-Pentamethyl-D-phenylalanyl-N¹—[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-    4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,β,β,3,5-Pentamethyl-D-phenylalanyl-N¹—[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β,3-Tetramethyl-L-phenylalanyl-N¹—[(1S,2E)-1-isopropyl-3-methyl-4-(4-methylpiperazin-1-yl)-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,    trifluoroacetic acid salt,-   N,    β,β,3,4-Pentamethyl-L-phenylalanyl-N¹—[(1S,2E)-1-isopropyl-3-methyl-4-(4-methylpiperazin-1-yl)-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β,3,5-Pentamethyl-L-phenylalanyl-N¹—[(1S,2E)-1-isopropyl-3-methyl-4-(4-methylpiperazin-1-yl)-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   1-{(2E,4S)-2,5-Dimethyl-4-[methyl(N,    β,β,3-tetramethyl-L-phenylalanyl-3-methyl-L-valyl    hex-2-enoyl}-D-prolyl-N-benzyl-D-prolinamide trifluoroacetic acid    salt,-   1-{(2E,4S)-2,5-Dimethyl-4-[methyl(N,    β,β,3,4-pentamethyl-L-phenylalanyl-3-methyl-L    -valyl)amino]hex-2-enoyl}-L-prolyl-N-benzyl-L-prolinamide,-   1-{(2E,4S)-2,5-Dimethyl-4-[methyl(N,    β,β,3,5-pentamethyl-L-phenylalanyl-3-methyl-L-    valyl)amino]hex-2-enoyl}-L-prolyl-N-benzyl-L-prolinamide,-   N,    β,β,3,4-Pentamethyl-L-phenylalanyl-N¹—[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β,3,4-Pentamethyl-D-phenylalanyl-N¹—[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β,3,5-Pentamethyl-L-phenylalanyl-N¹—[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1    -isopropyl-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β,β-Tetramethyl-L-phenylalanyl-N¹—[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   3-Chloro-N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β,3,4-Pentamethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β,3,4-Pentamethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-    -1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β,3,5-Pentamethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β,3,5-Pentamethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β,3-Tetramethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β,3-Tetramethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   3-Chloro-N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   3-Chloro-N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1    -yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   (E,4S)-4-[((2S)-3,3-dimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-N-hydroxy-2,5-dimethyl-2-hexenamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-(4-methyl-1-piperazinyl)-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamide,-   (2S)-N-[(1S,2E)-1-isopropyl-3-methyl-4-(4-morpholinyl)-4-oxo-2-butenyl]-N,3,3-trimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamide,-   1-{(2E,4S)-2,5-dimethyl-4-[methyl(N-methyl-3-phenyl-L-valyl-3-methyl-L-valyl)amino]hex-2-enoyl}-D-prolyl-N-benzyl-D-prolinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[(2S)-2-((1R,2R    )-1-methoxy-2-methyl-3-oxo-3-{[(1S)-2-phenyl-1-(1,3-thiazolyl-2-yl)ethyl]amino}propyl)pyrrolidin-1-yl]-3-methyl-4-oxobut-2-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-((1S,2E)-4-{(2S)-2-[(1R,2R)-1,3-dimethoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl}-1-isopropyl-3-methyl-4-oxobut-2-enyl)-N¹,3-dimethyl-L-valinamide,-   N-methyl-3-phenyl-L-valyl-N¹-[(1S,2E)-1-isopropyl-4-((2S)-2-{(1R,2R)-1-methoxy-2-methyl-3-oxo-[(2-phenethyl)amino]propyl}pyrrolidin-1-yl)-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-4-((2S)-2-{(1R,2R)-1-methoxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidin-1-yl)-3-methyl-4-oxovalinamide,-   Methyl N,    β,β-trimethyl-L-phenylalanyl-3-methyl-L-valyl-N-methyl-L-valyl-L-prolinate,-   N,    β,β-trimethyl-L-phenylalanyl-3-methyl-L-valyl-N-methyl-L-valyl-L-prolyl-N-benzyl-L-prolinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1    -isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(2-phenethoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-((1S,2E)-4-{4-[3-(dimethylamino)propyl]piperazin-1-yl}-1-isopropyl-3-methyl-4-oxobut-2-enyl)-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹—[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1-isopropyl    -methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S,4R)-2-(methoxycarbonyl)-4-hydroxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2R)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S,4R)-2-carboxy-4-hydroxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2R)-2-carboxypyrrolidin-1-yl]-1    -isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   3-cyclohexyl-N-methyl-L-valyl-N¹-{(1S,2E)-1-isopropyl-4-[(2S)-2-((1R,2R)-1-methoxy-    2-methyl-3-oxo-3-{[(1S)-2-phenyl-1-(1,3-thiazol-2-yl)ethyl]amino}propyl)pyrrolidin-1-yl]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   3-cyclohexyl-N-methyl-L-valyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   3-cyclohexyl-N-methyl-L-valyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxy-2,5-dihydro-1H-pyr    1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[(2S)-2-(methoxycarbonyl)piperidin-1-yl]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypiperidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   3-cyclohexyl-N-methyl-L-valyl-N¹-[(1S,2E)-1-isopropyl-4-((2S)-2-{(1R,2    2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidin-1-yl)-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[methoxy(methyl)-amino]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(1,1′-biphenyl-4-ylmethyl)amino]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,β,β-trimethyl-L-phenylalanyl-N¹-((1R,2E)-4-{[4-(benzyloxy)benzyl]oxy}-1-isopropyl-3-methyl-4-oxobut-2-enyl)-N¹,3-dimethyl-L-valinamide,-   (2E,4S)-4-[{N-[(2S)-2-(1-adamantyl)-2-(methylamino)ethanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoic    acid,-   (2E,4S)-4-[{N-[(2R)-2-(1-adamantyl)-2-(methylamino)ethanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoic    acid,-   (2E,4S)-2,5-dimethyl-4-(methyl{3-methyl-N-[(2S)-2-(methylamino)-2-phenylethanoyl]-L-    valyl}amino)-2-hexenoic acid,-   (2E,4S)-2,5-dimethyl-4-(methyl{3-methyl-N-[(2,2,4-trimethylthiomorpholin-3-yl)carbonyl]-L-valyl}amino)hex-2-enoic    acid,-   (2E,4S)-2,5-dimethyl-4-(methyl{3-methyl-N-[(2S)-2-(methylamino)-2-(1-methyl-1,2,3,4-tetrahydro-1-naphthalenyl)ethanoyl]-L-valyl}amino)-2-hexenoic    acid,-   (2E,4S)-2,5-dimethyl-4-(methyl{3-methyl-N-[(2R)-2-(methylamino)-2-(1-methyl-1,2,3,4-tetrahydro-1-naphthalenyl)ethanoyl]-L-valyl}amino)-2-hexenoic,-   3-hydroxy-N-methyl-L-valyl-N¹-[(1    S,2E)-4-ethoxy-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamide,-   3-hydroxy-N,N-dimethyl-L-valyl-N¹-[(1    S,2E)-4-ethoxy-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamide,-   1-methyl-L-prolyl-N¹-[(1    S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-va-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(ethoxysulfonyl)-1-isopropylprop-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-sulfoprop-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(ethoxysulfonyl)-1-isopropylbut-2-enyl]N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-sulfobut-2-enyl]N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2Z)-1-isopropyl-3-sulfobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(dimethoxyphosphoryl)-1-isopropylprop-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-3-[hydroxy(methoxy)phosphoryl]-1-isopropylprop-2-enyl}-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-phosphonoprop-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(diethoxyphosphoryl)-1-isopropyl-but-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-phosphonobut-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-4-(1,3-thiazol-2-yl)but-2-enyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-4-hydroxy-1-isopropyl-3-methyl-4-phenylbut-2-enyl}-N-1,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E,4R)-4-hydroxy-1-isopropyl-3-methyl-4-phenylbut-2-enyl}-N-1,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹[(1S,2E,4S)-4-hydroxy-1-isopropyl-3-methyl-4-phenylbut-2-enyl}-N-1,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹[(1S,2E)-4-hydroxy-1-isopropyl-3-methyl-4-(1,3-thiazol-2-yl)but-2-enyl]-N-1-,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-4-hydroxy-1-isopropyl-3-methylpent-2-enyl]-N-1,3-dimethyl-L-valinamide,-   6-{[3,3-Dimethyl-2-(3-methyl-2-methylamino-3-phenyl-butyrylamino)-butyryl]-methyl-amino}-2,4,7-trimethyl-octa-2,4-dienoic    acid,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-4-phenylbut-2-enyl]-N-1,3-dimethyl-L-valinamide,-   (2S)-N-[(1S,2E)-4-hydroxy-1-isopropyl-3-methyl-2-butenyl]-N,3,3-trimethyl-2-{[(S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamide,-   4-(dimethylsulfonio)-N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamide    trifluoroacetic acid,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-2-pentenyl]-N¹,3-dimethyl-L-valinamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹-{(1S)-1-[(Z)-(2,5-dioxo-4-imidazolidinylidene)methyl]-2-methylpropyl}-N¹,3-dimethyl-L-valinamide,-   (2S)-N-[(1S,2E)-1-isopropyl-3-(1H-tetraazol-5-yl)-2-propenyl]-N,3,3-trimethyl-2-{[(2S)-    3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamide,-   (2E,4S)-2,5-dimethyl-4-(methyl{3-methyl-N-[(2S)-3-methyl-2-(methylamino)-3-phenylbutyl]-L-valyl}amino)-2-hexenoic    acid,-   (2S)-N-[(1S,2E)-4-(1H-imidazol-2-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-    N,3,3-trimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamide,-   N,    β,β-trimethyl-L-phenylalanyl-N¹,3-dimethyl-N¹-{(1S)-2-methyl-1-[(Z)-(3-methyl-2,5    -dioxoimidazolidin-4-ylidene)methyl]propyl}-L-valinamide,-   N-{1-[(5-Hydroxy-1-isopropyl-3-methyl-4-oxo-pent-2-enyl)-methyl-carbamoyl]-2,2-dimethyl-propyl}-3-methyl-2-methylamino-3-phenyl-butyramide,-   Ethyl    (E,4S)-4-[((2S)-2-{[(2S)-2,3-dimethyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate,-   (E,4S)-4-[((2S)-2-{[(2S)-2,3-dimethyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoic    acid,-   Ethyl    (E,4S)-4-[((2S)-2-{[(2S)-2-azido-3-methyl-3-phenylbutanoyl]amino}-3,3    -dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate,-   Ethyl    (E,4S)-4-[((2S)-2-{[(2R)-2-azido-3-methyl-3-phenylbutanoyl]amino}-3,3    -dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate,-   Ethyl    (E,4S)-4-[((2S)-2-{[(2S)-2-azido-3-methyl-3-phenylbutanoyl]amino}-3,3    -dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate,-   Ethyl    (E,4S)-4-[((2S)-2-{[(2R)-2-azido-3-methyl-3-phenylbutanoyl]amino}-3,3    -dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate,-   Ethyl    (E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-(benzyl)-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate,-   (E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-benzyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoic    acid,-   Ethyl    (E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-allyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate,-   (E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-allyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoic    acid,-   Ethyl    (E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-ethyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate,-   (E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-ethyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoic    acid,-   Ethyl    (E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-methylsulfanyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate,-   (E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-methylsulfanyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoic    acid,-   (E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-methylsulfonyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoic    acid,-   Ethyl    (E,4S)-4-[{(2S)-2-[(2-methoxy-3-methyl-3-phenylbutanoyl)amino]-3,3-dimethylbutanoyl}(methyl)amino]-2,5-dimethyl-2-hexenoate,-   (E,4S)-4-[{N-[(2S)-2-methoxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoic    acid,-   (E,4S)-4-[{N-[(2R)-2-methoxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoic    acid,-   (E,4S)-4-[{N-[(2S)-2-hydroxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoic    acid,-   (E,4S)-4-[{N-[(2R)-2-hydroxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoic    acid and-   (2S)-N-[(1S,2E)-4-hydrazino-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N,3,3-trimethyl    -2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamide.

DETAILED DESCRIPTION OF THE INVENTION

Compounds of this invention may be prepared as outlined in the followingschemes.

As described in Scheme 1,2-oxopropanoic acid 1 is reacted with methyliodide in a solvent mixture of aqueous sodium hydroxide-tetrahydrofuran(THF) to afford pyruvic acid 2 which is further converted to acid 3 bysequential treatment with trimethylsilyldiazomethane (TMSCHN₂), sodiumborohydride(NaBH₄) and lithium hydroxide.

As further described in Scheme 2, ester 4 is reacted with methyl iodidein the presence of silver oxide in ether to give ether 5 which is thenhydrolyzed with lithium hydroxide in a solvent mixture of THF-methylalcohol-water to give propionic acid 6.

Described in Scheme 3, reaction of propionic acid 7 with 1.1 equivalentof disulfide RSSR in the presence of lithium diisopropylamide (LDA) and1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU) affordssulfide 8 which is further oxidized with peroxyacetic acid to givesulfone 9.

According to Scheme 4, alkylation of amide 10 with R₂X, wherenon-limiting examples include methyl, benzyl, allyl (CH₂═CH—CH₂—) andazido, in the presence of sodium or potassium1,1,1,3,3,3-hexamethyldisilazane (Na(K)HMDS) gives alkylated product 11.Reaction of alkylated product 11 with lithium peroxide affords propionicacid 12.

As described in Scheme 5, tetralone 13 is reacted with isonitrile 14 inthe presence of potassium tert-butoxide to give ester 15 which isfurther reacted with hydrochloric acid then alkylated with methyl iodidein the presence of sodium hydroxide and hydrolyzed with sodium hydroxideto give carboxylic acid 16. Carboxylic acid 16 is reacted withmethylamine in the presence of borane-pyridine complex to give amine 17.

According to Scheme 6, reaction of carboxylic acid 18 with methylamineaffords substituted-amino acid 19.

According to Scheme 7, reaction of carboxylic acid 20 with formic acidand formalin affords substituted amine 21.

Described in Scheme 8, reaction of carboxylic acid 22 where E and A arehereinbefore defined with amine 23 in the presence of a coupling agentselected from 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (EDC) orbenzotriazole-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate(PyBOP) gives carboxylic acid 24.

As described in Scheme 9, reaction of carboxylic acid 25 with an amineNHRR in the presence of a coupling agent selected from1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) orbenzotriazole-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate(PyBOP) affords amide 26. Further, reaction of carboxylic acid 25 withan alcohol ROH in the presence of a coupling agent selected from1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) orbenzotriazole-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate(PyBOP) affords ester 27.

Described in Scheme 10, is the reaction of aldehyde 28 with(carboethoxyethylidene)triphenylphosphorane followed by reaction withlithium hydroxide to give carboxylic acid 29.

As described in Scheme 11, reaction of carboxylic 30 withN-methyl-N-methoxyamine in the presence of a coupling agent selectedfrom 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC)or benzotriazole-1-yl-oxy-tris-pyrrolidinophosphoniumhexafluorophosphate (PyBOP) affords amide 31 which is reduced withlithium aluminum hydride (LAH) to give aldehyde 33. Reaction of amide 31with RM, where M is a metal such as lithium, potassium, sodium ormagnesium affords ketone 32. Further reaction of ketone 32 with ahydride reducing agent selected from LAH or sodium borohydride givesalcohol 34 which may also be prepared by reaction of aldehyde 33 withRM. Reaction of acid 35 with1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC)followed by treatment with sodium borohydride gives alcohol 36 which canbe oxidized to aldehyde 33 by treatment with dimethylsulfoxide/oxalylchloride or manganese dioxide.

As described in Scheme 12, carboxylic acid is ozonized in methyl alcoholfollowed by treatment with dimethylsulfide to give in situ aldehyde 37.A 1,1-bisphosphonate ester 38 is treated with n-butyllithium followed byaldehyde 37 to give phosphonate 39 which is further reacted withtrimethylsilylbromide to give phosphonate 40. Aphosphorylmethylenesulfonate 41 is treated with n-butyl lithium followedby aldehyde 37 to afford sulfonate ester 42 which is reacted withtetrabutylammonium iodide to give sulfonic acid 43. Additionally,treating aldehyde 37 with phosphonate 44 affords hydantoin 45.

As described in Scheme 13, amide 31 is reacted with 47 (the lithium saltof trimethylsilyl ether 46) (which is prepared by reaction oftributyltin reagent 46 with n-butyl lithium) to give ester 48 which isthen treated with trifluoroacetic acid (TFA) to give carboxylic acid 49.

Described in Scheme 14, aldehyde 50 is reacted with(triphenylphosphoranylidene)acetonitrile 51 to give nitrile 52 which istreated with trifluoroacetic acid (TFA) to give amine 53. Reaction of 53with carboxylic acid 54 in the presence ofbenzotriazole-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate(PyBOP) affords nitrile 55. Treatment of nitrile 55 with trifluoroaceticacid (TFA) gives amine 56. Treatment of amine 56 with carboxylic acid 57in the presence ofO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU), 1-Hydroxy-7-azabenzotriazole (HOAt) anddiisopropylethylamine (DIEA)gives nitrile 58 which is further reactedwith trimethylsilylazide and dibutyltin oxide followed bytrifluoroacetic acid (TFA) to give tetrazole 59.

Described in Scheme 15, reaction of carboxylic acid 60 with amine 61 inthe presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (EDC) followed by treatment with trifluoroacetic acid(TFA) affords amine 62 which is reacted with carboxylic acid 63 in thepresence of benzotriazole-1-yl-oxy-tris-pyrrolidinophosphoniumhexafluorophosphate (PyBOP) followed by treatment with trifluoroaceticacid to give amine 64. Further reaction of amine 64 with carboxylic acid65 in the presence of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (EDC) followed by treatment with trifluoroacetic acid(TFA) affords proline derivative 66.

According to Scheme 16, imidazole 67 is reacted with chloromethyl ethylether in the presence of sodium hydride to give ether 68 which isreacted with n-butyllithium to afford lithium salt 69. Reaction of ester70 with lithium hydroxide followed by treatment withN,O-dimethylhydroxylamine in the presence of1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC)affords amide 71 which is reacted with lithium salt 69 to give imidazole72. Acid hydrolysis of imidazole 72 with HCl affords amine 73. Furtherreaction of amine 73 with carboxylic acid 60 in the presence of3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT) givesimidazole 74 which is further reacted with amino acid 75 in the presenceof DEPBT followed by treatment with trifluoroacetic acid to giveimidazole 76.

As described in Scheme 17, reaction of carboxylic acid 57 withmorpholine in the presence ofbenzotriazole-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate(PyBOP) affords amide 77 which is reduced with lithium aluminum hydrideto give aldehyde 78 which is coupled with amine 79 by reductivealkylation in the presence of zinc chloride and sodium cyanoborohydridefollowed by sequential treatment with lithium hydroxide (LiOH) andhydrochloric acid (HCl) acid to give 80.

Reactions are performed in a solvent appropriate to the reagents andmaterials employed and suitable for the transformation being effected.It is understood by those skilled in the art of organic synthesis thatthe various functionalities present on the molecule must be consistentwith the chemical transformations proposed. This may necessitatejudgement as to the order of synthetic steps, protecting groups, ifrequired, and deprotection conditions. Substituents on the startingmaterials may be incompatible with some of the reaction conditions. Suchrestrictions to the substituents which are compatible with the reactionconditions will be apparent to one skilled in the art.

Some of the compounds of the hereinbefore described schemes have centersof asymmetry. The compounds may, therefore, exist in at least two andoften more stereoisomeric forms. The present invention encompasses allstereoisomers of the compounds whether free from other stereoisomers oradmixed with other stereoisomers in any proportion and thus includes,for instance, racemic mixture of enantiomers as well as thediastereomeric mixture of isomers. The absolute configuration of anycompound may be determined by conventional X-ray crystallography.

According to a further aspect of the present invention there is provideda series of compounds of formula (I) or the pharmaceutically acceptablesalts thereof as hereinbefore defined for use in a method of treatmentof human or animal disease.

In particular, compounds of the invention are useful for the treatmentof cancer and cancerous tumors.

Further, compounds of the present invention inhibit tubulinpolymerization and are therefore are useful in the treatment of cancer.

In particular, the present invention relates to a method of treating orinhibiting the growth of cancerous tumors in a mammal with inherent oracquired resistance to chemotherapeutic agents used in chemotherapytreatment and in particular antimitotic agents by administering aneffective amount of a compound of formula (I) and pharmaceuticallyacceptable salts thereof.

Accordingly, it is one of the purposes of this invention to overcome theabove described limitations in cancer treatment by providing a methodfor treating tumors that are resistant to currently marketed antimitoticagents.

Further, compounds of the invention provide a method for the treatmentor prevention of cancerous tumors that express multiple drug resistance(MDR) or are resistant because of MDR in a mammal in need thereof.

Additionally, the present invention provides a method of treating,inhibiting the growth of, or eradicating a tumor in a mammal in needthereof wherein said tumor is resistant to at least one chemotherapeuticagent which method comprises providing to said mammal an effectiveamount of a compound of Formula (I).

Cytotoxicity Standard Pharmacological Test Procedures

Tubulin Polymerization Assays. For in vitro tubulin polymerizationassays, bovine microtubule-associated protein (MAP)-rich tubulin, PEMbuffer [80 mM Na-PIPES (pH 6.9), 1 mM MgCl₂, and 1 mM EGTA], and GTP arepurchased from Cytoskeleton (Denver, Colo.). MAP-rich tubulin (finalconcentration 1.5 mg/ml) is dissolved in cold PEM buffer containing 1 mMGTP (GPEM) and centrifuged at 12,000×g for 10 min at 4° C. The tubulinsolution (100 μl/well) is added rapidly to wells of a low-volume,96-well plate already containing duplicate aliquots (10 μl) of testcompounds in GPEM. Final compound concentrations are 0.1 or 1 μM.Control wells contained the same final concentration of DMSO (0.3%).After initiation of the reaction, absorbance at 340 nm is measured everyminute for 60 min at 24° C. using a SpectraMax Plus plate reader(Molecular Devices, Sunnyvale, Calif.). The resultant data is displayedin Table 1.

Biological Testing Results

TABLE 1 Tubulin Ex. No. KB* KB85* KBV1* Colo205* Polym.** 1 19.5 56 151479 2 34 62 162 37 3 70(2) 176(2) 1758(2) 51 4 375 536 42% 61 5 207 45940% 57 6 64(2) 123(2)  33%(2) 64 7 47 68 2188 32 8 35 56 1622 68 91.4(3)   23(3) 21% 56 10 1.5(2)  19.7(2)  2138(2) 42 11 0.56 24  8% 4112 48 65 1334 51 13 172(2)  352(2) 1771(2) 11 14 45 135 1647 51(5) 157.2 18 354 77 16 39 60 803 62 17 1.9 15 363 71 18 74 519  6% 76 19 5.950 1552 66 20 82 664  3% 80 21 1.6 6.2 275 89 22 70 599 12% 95 23 2.56.2 362 88 24 62 180 17% 94 25 39(2) 227(2)  7% 20 75 26 33(2) 60(2)2326(2) 71(2) 27 43.5(6)   73.5(6)  1301(6) 23 69 28 1.07(3)   21.9(3)  14%(3) 0.25 56(3) 29 0.75(2)    4.1(2)   180(2) 62 30 3.3(2)   6(2) 96(2) 45 31 0.9 5.1 191 69 32 12 45 636 99(2) 33 18 53 1533 19 340.25(2)    4.2(2)    5%/1948 64 35 1.5(3)   7.3(3)   291(3) 82 36 62 205 0% 97 37 6.4 21 1643 86 38 20 529  0% 75 39 15(2)  22(2)  366(2) 29 403.5 116  4% 93*** 41 16 51 1099 96 42 184 1850  0% 96 43 515 1577  0% 9144 1.5 6.2 475 83 45 3.7 16 493 62(2) 46 450 1719  1% 99 47 3.5 17 27288(2) 48 58 184 15% 91(2) 49 5.2 17 150 62(2) 50 64 270  9% 84(2) 51 2261 1772 82 52 11 47 392 86 53 667 1675 29% 11 54 136 846  8% 55 55 603646 13% 66 56 468 575 11% 57 1756(2)  2610(2)    7%(2) 75 58 427 550 15%89(2) 59 5.5 6.6 76 93 60 9.1 17 427 94 61 151 293 >3000 40*** 622952(2)  26%(2)   10%(2) 65(2) 63 1% 0%  0% 42 64 2% 1%  0% 86 65 11% 1%  0% 44 66 25%  0%  0% 91 67 0% 0%  0% 21 68 1976(2)  2077(2)    8%(2)45 69 0% 0%   3% 59 70 0% 0%   3% 69 71 1304 1751   0% 64 72 0% 0%  4%53 73 582 565 1787 33 74 19 54 466 73 75 124 186 632 83*** 76 16 22 19063*** 77 13 19 364 97 78 63 172 1455 75 79 27 53 1582 90*** 80 725 13261805 70 81 454(2)  447(2) 1594(2) 71 82 713(6)  1330(6)  1938 > 3000(6)  5.1(16) 51/47 83    124/>3000    215/>3000 2398 > 3000   46 0 84403(2)  664(2) 2358(2) 180 69 85 1072 1738 10% 59 86 1148 1819  5% 71/7131 87 104(2)  312(2)  14%(2) 28 88 218 1364  0% 91 89 5.1 9.9 230 95 9043 68 1134 76*** 91 21%  >3000 >3000 >1000 84**** 92 861(2)  990(2) 30% 53/210 94 93 2511 30%  30% — 94 1297(2)  1332(2)   30%(2) 460 73 95 575756 15% >1000 — 96 846(3)  1122(3)   11%(2)/2630   340 46 97 1910 332017%@6000 >1000 — 98 1151 1205 42%@6000 270 63 99 3476 363816%@6000 >1000 — 100 957 1151 5260 190 54 101 1100 1824 4580 >1000 — 102693 1151 40%@6000 280 79 103 1749(2)  1659(2)/30%   5% 760 80 1041731/3000 2126/3000 2398/3000 >1000 38 105  43%/2691 21%(2)    0%(2)1738(2) 64 106 1416 1229 22% 930 54 107 >3000 >3000 >3000 >1000 21108 >3000 >3000 >3000 >1000 21 109 65(4) 227(4) 2077(3)/23%    110173/2512/ 263/26%/ 22%/23% >1000 19 2630/7% 22%/10% %/24%@ @3000/5%@3000/0% 3000/4% @3000 @3000 @3000 Data is the average of the number ofdeterminations in parenthesis. *IC50 in nM or % inhibition @ 3000 nM **%inhibition at 0.3 uM ***% inhibition at 0.5 uM ****% inhibition at 2 uM

Cell Survival Assay: Standard Pharmacological Test Procedure. Theconcentration of candidate inhibitor required to inhibit 50% of cellgrowth (IC₅₀) is done according to previously reported methods.(Discafani, C. M., Carroll, M. L., Floyd Jr., M. B. F., Hollander, I.J., Husain, Z., Johnson, B. D., Kitchen, D., May, M. K., Malo, M. S.,Minnick Jr., A. A., Nilakantan, R., Shen, R., Wang Y-F., Wissner, A.,Greenberger, L. M. Irreversible inhibition of epidermal growth factorreceptor tyrosine kinase with in vivo activity byN-[4-[3-bromophenyl)amino]-6-quinaxolinyl]-2-butynamide (CL-387,785),Biochem. Pharmacol. 57: 917-925,1999) Briefly, cells are plated in 100μl of media in the morning of day 1 and allowed to adhere to the platesfor 2-6 hr. Compounds are serially diluted into media as 2× stocks and100 μl added to cells in duplicate. Compounds are incubated with cellsfor 3 days. At the end of the incubation period the sulforhodamine B(SRB) assay, which measures protein content as an assessment of cellsurvival, is performed as described previously (Skehan P, Storeng R,Scudiero D, Monks A, McMahon J, Vistica D, Warren JT, Bokesch H, KenneyS, Boyd MR. New colorimetric cytotoxicity assay for anticancer-drugscreening. J Natl Cancer Inst 1990 Jul. 4;82(13):1107-12) with somemodification as follows. For KB cells with 20% FCS, media is gentlydecanted and replaced with 200 μl of serum-free media mixed with cold50% TCA for a final concentration of 10% TCA. The plates are incubatedfor 1 hr at 4° C., followed by washing 5 times in cold distilled water,then air dried overnight. The fixed cells are strained for 10 min with80 μl of 0.04% SRB solution prepared in 1% glacial acetic acid. Stain isdiscarded and plates washed 5 times in 1% glacial acetic acid, thenair-dried until completely dry. Stained cell product is dissolved in 150μl of 10 mM Trizma Base and placed on a shaker for 20 minutes untilfully dissolved. Absorbance is read on a Victor V multi-label platereader (Perkin Elmer, Gaithersburg, Md.). The resultant data isdisplayed in Tables 1 and 2.

Results of Standard Pharmacological Test procedure

TABLE 2 Ex. No. HCT-15 LOX S1-M1 S1 A375 KM20 NCl-H1299 92 1380 1072 1202  27  26(2)  50(4) 336(2) 657(2)  38 105 1738  82 1499(2) 630(4)663(4) 4.6(4) 1572(6) 84 1259 550 832 87  759 151 295 26  46 109 19 3511 9 110     19% Data is the average of the number of determinations inparenthesis. IC50 in nM or % inhibition @ 3000 nM

Growth of Tumors in Nude Mice. Drug efficacy studies in mice areperformed similar to previously reported studies. (Discafani, C. M.,Carroll, M. L., Floyd Jr., M. B. F., Hollander, I. J., Husain, Z.,Johnson, B. D., Kitchen, D., May, M. K., Malo, M. S., Minnick Jr., A.A., Nilakantan, R., Shen, R., Wang Y-F., Wissner, A., Greenberger, L. M.Irreversible inhibition of epidermal growth factor receptor tyrosinekinase with in vivo activity byN-[4-[3-bromophenyl)amino]-6-quinaxolinyl]-2-butynamide (CL-387,785),Biochem. Pharmacol. 57: 917-925, 1999) Briefly, athymic nu/nu femalemice (Charles River Laboratories) are implanted SC (subcutaneously) with1.5×10⁶ LOX melanoma cells. When tumors attain a mass of between 80 and120 mg (day 0), animals are randomized into treatment groups eachcontaining either 5 or 10 animals, (dependent upon the experiment). Insome experiments, tumors are allowed to grow up to 2.5 grams in sizebefore drug treatment is initiated. After staging, animals are treatedintravenously (IV) with one or more doses of test compound formulated insaline, or vehicle control. The doses chosen for both paclitaxel andvincristine are between 80 and 90% of the maximum tolerated dose foreach drug. Tumor mass ([length×Width ²]/2) is determined once a week forup to 35 days. The relative tumor growth (mean tumor mass on daymeasured divided by the mean tumor mass on day zero) and the percentTumor/Control (%T/C) are then calculated for each treatment group forthe duration of each experiment. The %T/C is defined as the MeanRelative Tumor Growth of the Treated Group divided by the Mean RelativeTumor Growth of Vehicle Control Group multiplied by 100. The data areanalyzed via a one-sided Student's t-test. A p-value ≦0.05 indicates astatistically significant reduction in relative tumor growth of treatedgroup compared to that of the vehicle control group. The results aredisplayed in Table 3.

Effect on the Growth of the Human Melanoma Carcinoma Lox in a XenograftModel

TABLE 3 Ex. Dose % T/C on day No. (mg/kg) Route 7 9 10 12 13 14 27 10 IV13 2 27 10 IV 11 2 28 10 IV 10 4 109 3 IV 24 8 35 2 IV 5 1 29 10 IV 14 926 15 IV 4 30 5 IV 11 3 Groups of 5 female nu/nu mice are injected with1.5 × 10⁶ Lox cells. Animals bearing staged tumors are treated IV or IPwith vehicle, Examples, paclitaxel or vincristine at the doses indicatedon days 1, 5, 9. Relative tumor growth is determined during theexperiment and % T/C calculated. Statistical analyses are Student'st-test of treated time points vs. vehicle (P < 0.01)

The compounds of Formula (I) may be obtained as inorganic or organicsalts using methods known to those skilled in the art (Richard C.Larock, Comprehensive Organic Transformations, VCH publishers, 411-415,1989). It is well known to one skilled in the art that an appropriatesalt form is chosen based on physical and chemical stability,flowability, hydroscopicity and solubility.

Pharmaceutically acceptable salts of the compounds of Formula (I) withan acidic moiety may be formed from organic and inorganic bases. Forexample with alkali metals or alkaline earth metals such as sodium,potassium, lithium, calcium, or magnesium or organic bases andN-tetraalkylammonium salts such as N-tetrabutylammonium salts.Similarly, when a compound of this invention contains a basic moiety,salts may be formed from organic and inorganic acids. For example saltsmay be formed from acetic, propionic, lactic, citric, tartaric,succinic, fumaric, maleic, malonic, mandelic, malic, phthalic,hydrochloric, hydrobromic, phosphoric, nitric, sulfuric,methanesulfonic, naphthalenesulfonic, benzenesulfonic, toluenesulfonic,camphorsulfonic, and similarly known acceptable acids. An additionalacid is trifluoroacetic acid (TFA). The compounds can also be used inthe form of esters, carbamates and other conventional prodrug forms,which when providing in such form, convert to the active moiety in vivo.

Providing, means to make available and is intended to include directadministration as well as in vivo (e.g. pro-drugs.) of compounds used inthe method of treating, inhibiting the growth of, or eradicating a tumorin a mammal in need thereof wherein said tumor is resistant to at leastone chemotherapeutic agent which method comprises providing to saidmammal an effective amount of a compound of Formula (I).

Based on the results of these standard pharmacological test procedures,the compounds of this invention are useful as agents for treating,inhibiting the growth of or eradicating tumors resistant tochemotherapeutic agents and in particular to antimitotic compounds,including paclitaxel. It is estimated that a 70 kg human will receive anexposure, or AUC, to drug that is nearly equivalent to the efficaciousdose in mice upon a preferred effective intravenous dose of about 0.6 toabout 1.3 mg. Therefore, a more preferred effective regimen for optimumresults would be from about 0.8 to about 200 micrograms (ug)/kg of bodyweight per cycle and such dosage units are employed that a total of fromabout 0.05 mg to about 150 mg of the active compound for a subject ofabout 70 kg of body weight are administered intravenously in a cycle. Acycle may be once every 1, 2 or 3 weeks as typically used for otheranti-microtubule drugs in the clinic.

Tumors are selected from the group consisting of breast, colon, lung,prostate, melanoma, epidermal, leukemia, kidney, bladder, mouth, larynx,esophagus, stomach, ovary, pancreas, liver, skin and brain.

The dosage regimen for treating mammals may be adjusted to provide theoptimum therapeutic response. For example, several divided doses may beprovided per cycle or the dose may be proportionally reduced asindicated by the exigencies of the therapeutic situation. A decidedlypractical advantage is that these active compounds may be provided inany convenient manner such as by the oral, intravenous, intramuscular orsubcutaneous routes. Preferably compounds of the invention are providedby intravenous routes.

The active compounds may be orally provided, for example, with an inertdiluent or with an assimilable edible carrier, or they may be enclosedin hard or soft shell gelatin capsules, or they may be compressed intotablets or they may be incorporated directly with the food of the diet.For oral therapeutic administration, these active compounds may beincorporated with excipients and used in the form of ingestible tablets,buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafersand the like. Such compositions and preparations should contain at least0.1% of active compound. The percentage of the compositions andpreparations may, of course, be varied and may conveniently be betweenabout 2% to about 60% of the weight of the unit. The amount of activecompound in such therapeutically useful compositions is such that asuitable dosage will be obtained. Preferred compositions or preparationsaccording to the present invention are prepared so that an oral dosageunit form contains between 0.5 and 1500 mg of active compound. Thetablets, troches, pills, capsules and the like may also contain thefollowing: a binder such as gum tragacanth, acacia, corn starch orgelatin; excipients such as dicalcium phosphate; a disintegrating agentsuch as corn starch, potato starch, alginic acid and the like; alubricant such as magnesium stearate; and a sweetening agent such assucrose, lactose, or saccharin may be added or a flavoring agnet such aspeppermint, oil of wintergreen or cherry flavoring. When the dosage unitform is a capsule, it may contain, in addition to materials of the abovetype, a liquid carrier. Various other materials may be present ascoatings or to otherwise modify the physical form of the dosage unit.For instance, tablets, pills or capsules may be coated with shellac,sugar or both. A syrup or elixir may contain the active compound,sucrose, as a sweetening agent, methyl and propylparabens aspreservatives, a dye and flavoring such as cherry or orange flavor. Ofcourse, any material used in preparing any dosage unit form should bepharmaceutically pure and substantially non-toxic in the amounts used.In addition, these active compounds may be incorporated intosustained-release preparations and formulations.

The active compounds of the invention may also be provided parenterallyor intraperitoneally. Solutions or suspensions of these active compoundsas a free base or pharmacologically acceptable salt can be prepared inwater suitably mixed with a surfactant such as hydroxypropylcellulose.Dispersions can also be prepared in glycerol, liquid polyethyleneglycols, and mixtures therof in oils. Under ordinary conditions ofstorage and use, these preparations contain a preservative to preventthe growth or microorganisms.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. In all cases, the form must be sterile and must be fluid tothe extent that easy syringability exists. It must be stable under theconditions of manufacture and starage and must be prepared against thecontaminating action of microorganisms such as bacteria and fungi. Thecarrier can be a solvent or dispersion medium containing, for example,water, ethanol, polyol (e.g., glycerol, propylene glycol and liquidpoly-ethylene glycol), suitable mixtures thereof, and vegetable oils.Preferably, compounds of this invention are provided by an IV route.

The compounds of this invention and their preparation are illustrated bythe following non-limiting examples.

General Procedure I (Amide Formation)

Method A: To a cooled (0° C., ice-water bath) solution of the carboxylicacid (1 mmol) in anhydrous solvent (acetonitrile or dimethylformamide,3-5 ml) are added hydroxybenzotriazole (1.1 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.2 mmol)under an inert atmosphere. The cooling bath is removed, and theresulting mixture is stirred at room temperature for 2-15 hours, until aclear solution is obtained. To this mixture is added a solution of anamine in anhydrous solvent (3-5 ml) at 0° C., and the resulting mixtureis stirred at room temperature for 2-24 hours until Mass spectraanalysis indicated no starting material left in the reaction mixture.The reaction mixture is concentrated in vacuo, and the product ispurified by separation using preparative HPLC.

Alternatively, the product is isolated by aqueous work-up procedure: thereaction mixture is diluted with ethyl acetate (100 ml/mmol), and theorganic layer is washed with saturated aqueous sodium bicarbonatesolution and saturated sodium chloride solution, dried over anhydroussodium sulfate, and concentrated in vacuo. The product is purified bychromatography (silica gel).

Method B: ToN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(0.1 mmol, Andersen, R. WO 99/32509) in acetonitrile (5.0 mL) at 25° C.is added 1-hydroxybenzotriazole hydrate (0.12 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.33 mmol).After 2-18 hours an amine (0.4 mmol) is added. After 1-18 hours thereaction mixture is concentrated in vacuo, dissolved indimethylformamide, and the product is purified by reverse phase HPLC(0.01% aqueous trifluoroacetic acid/acetonitrile gradient system).

Method C: A mixture ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(1.0 equivalent, Andersen, R. WO 99/32509), hydroxybenzotriazole hydrate(1.2 equivalents), 1-(3-dimethylaminopropyl)-3-ethylcarbodiiminehydrochloride (1.2 equivalents), an amine (1.2 equivalents) and Hunig'sbase (2.0 equivalents) in anhydrous dimethylformamide (approx 6 mmol)are stirred under a nitrogen atmosphere overnight. The solvent isremoved, the residue is taken up in methanol, and the product ispurified by reverse phase HPLC (0.01% trifluoroacetic acid inwater/acetonitrile).

General Procedure II (Ester Hydrolysis):

A peptide ester is dissolved in methanol (24 mmol), and cooled to 0° C.(ice-water bath). To this solution is added water (8 mmol) and aqueouslithium hydroxide solution (8 mol equivalents). The cooling bath isremoved, and the resulting mixture is stirred at room temperature for 15hours. Methanol is removed in vacuo, and the residual aqueous mixture iscooled with an ice water bath, and acidified to pH 5.5-6.0 with aqueous1N citric acid solution. The precipitate is collect by filtration, andthe solid is washed with cold water, and dried over high vacuum.Alternatively, the product can be purified by preparative reverse phaseHPLC.

EXAMPLE 1N,β,β,3-Tetramethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure to that described in General Procedure IMethod A,N,β,β,3-tetramethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(100 mg, 0.205 mmol) is treated with hydroxybenzotriazole (36 mg, 0.27mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(59 mg, 0.31 mmol) in anhydrous acetonitrile (3 ml) at room temperaturefor 15 hours, followed by addition of morpholine (0.036 ml, 0.41 mmol).After stirring at room temperature for 15 hours, the product is purifiedusing the aqueous work-up method, to provide the title compound (89 mg,78%) as a orange amorphous solid.

MS (ES): m/z 557.4 (M+H). Analytical HPLC: (4.6×150 mm YMC Pro Pack C18column eluted with 10 to 100% acetonitrile in water containing 0.02%formic acid over 29 minutes): 92.55% (at 17.3 minutes) ofN,β,β,3-tetramethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,and 5.99% (at 18.8 minutes) of another diastereomer.

EXAMPLE 2N,β,β,3,4-Pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3,4-pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(107.7 mg, 0.215 mmol) is treated with hydroxybenzotriazole (44 mg,0.323 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimidehydrochloride (65 mg, 0.344 mmol) in anhydrous dimethylformamide (3 ml)at room temperature for 2 hours, followed by addition of morpholine(0.038 ml,0.43 mmol). After stirring at room temperature for 36 hours,the product is purified by preparative HPLC, to provide thetrifluoroacetic acid salt of the title compound (82 mg, 66.8%) as awhite amorphous solid MS (ES): m/z 571.4 (M+H). IR cm⁻¹:2966,1669,1634.Analytical HPLC: (4.6×150 mm Prodigy ODS(3) 320486 column eluted with 10to 100% acetonitrile in water containing 0.02% formic acid over 30minutes):96.015% (at 16.506 minutes) ofN,β,β,3,4-pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,and 0.78% (at 16.923 minutes), 0.747% (at 17.441 minutes), and 0.29% (at17.71 minutes) of three other diastereomers.

EXAMPLE 3N,β,β,3,5-Pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3,5-pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(104 mg, 0.2073 mmol) is treated with hydroxybenzotriazole (45 mg, 0.333mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(65 mg, 0.344 mmol) in anhydrous dimethylformamide (3 ml) at roomtemperature for 2 hours, followed by addition of morpholine (0.04ml,0.458 mmol). After stirring at room temperature for 36 hours, theproduct is purified by preparative HPLC to provide the trifluoroaceticacid salt of the title compound (90.0 mg, 87.1%) as a white amorphoussolid. MS (ES): m/z 571.4 (M+H). Analytical HPLC: (4.6×150 mm YMC PackPro C18 column eluted with 10 to 100% acetonitrile in water containing0.02% formic acid over 35 minutes): 97.39% (at 18.878 minutes) ofN,β,β,3,5-pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,and 0.66% (at 20.395 minutes) of the other diastereomer.

EXAMPLE 4N,β,β,3,4-Pentamethyl-D-phenylalanyl-N¹[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3,4-pentamethyl-D-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(150 mg, 0.299 mmol) is treated with hydroxybenzotriazole (60 mg, 0.444mmol) and 1-(3-dimethylaminopropyl)-3ethylcarbodiimide hydrochloride (86mg, 0.449 mmol) in anhydrous acetonitrile (3 ml) at room temperature for2 hours, followed by addition of morpholine (0.052 ml, 0.596 mmol).After stirring at room temperature for 36 hours, the product is purifiedby preparative HPLC, to provide the trifluoroacetic acid salt of thetitle compound (60 mg, 35.2%) as a white amorphous solid. MS (ES): m/z571.4 (M+H). IR cm^(−1:)2964, 1669, 1636. Analytical HPLC: (4.6×150 mmYMC Pack Pro C18 column eluted with 10 to 100% acetonitrile in watercontaining 0.02% formic acid over 35 minutes):98.34% (at 13.604 minutes)ofN,β,β,3,4-pentamethyl-D-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,and 0.28% (at 13.3 minutes) and 0.26% (at 14.4 minutes) of two otherdiastereomers.

EXAMPLE 5N,β,β,3,5-Pentamethyl-D-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3,5-pentamethyl-D-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(150 mg, 0.299 mmol) is treated with hydroxybenzotriazole (60 mg, 0.444mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(86 mg, 0.449 mmol) in anhydrous acetonitrile (3 ml) at room temperaturefor 2 hours, followed by addition of morpholine (0.052 ml, 0.596 mmol).After stirring at room temperature for 36 hours, the product is purifiedby preparative HPLC, to provide the trifluoroacetic acid salt of thetitle compound (100 mg, 58.6%) as a white amorphous solid. MS (ES): m/z571.4 (M+H). IR cm⁻¹: 2965, 1669, 1636. Analytical HPLC: (4.6×150 mm YMCPack Pro C18 column eluted with 10 to 100% acetonitrile in watercontaining 0.02% formic acid over 35 minutes): 86.47% (at 13.847minutes) ofN,β,β,3,5-pentamethyl-D-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-morpholin-4-yl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,and 10.04% (at 13.448 minutes) of the other diastereomer.

EXAMPLE 6N,β,β,3-Tetramethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-(4-methylpiperazin-1-yl)-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamidetrifluoroacetic acid salt

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3-tetramethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(100 mg, 0.205 mmol) is treated with hydroxybenzotriazole (36 mg, 0.27mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(59 mg, 0.31 mmol) in anhydrous acetonitrile (10 ml) at room temperaturefor 15 hours, followed by addition of N,N-diiospropylethylamine (0.143ml, 0.82 mmol) and 1-methylpiperazine (0.045 ml, 0.41 mmol). Afterstirring at room temperature for 4 hours, the product is purified bypreparative HPLC, to provide the trifluoroacetic acid salt of the titlecompound (132 mg, 94%) as a white amorphous solid. MS (ES): m/z 570.4(M+H). IR cm^(−1:) 2969.14, 1675.70, 1629.86, 1203.89, 1134.29.Analytical HPLC: (4.6×150 mm YMC Pack Pro C18 column eluted with 10 to100% acetonitrile in water containing 0.02% formic acid over 35minutes): 92.03% (at 7.5 minutes) ofN,β,β,3-tetramethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-(4-methylpiperazin-1-yl)-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,and 4.87% (at 9.2 minutes) of the other diastereomer.

EXAMPLE 7N,β,β,3,4-Pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-(4-methylpiperazin-1-yl)-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3,4-pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(105 mg, 0.2093 mmol) is treated with hydroxybenzotriazole (44 mg, 0.326mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(65 mg, 0.339 mmol) in anhydrous dimethylformamide (3 ml) at roomtemperature for 3 hours, followed by addition of 1-methylpiperazine(0.032 ml, 0.32 mmol). After stirring at room temperature for 15 hours,the product (108 mg, 87.7%) is purified by preparative HPLC, to providethe trifluoroacetic acid salt of the title compound (66.1 mg, 53.7%) asa white solid. MS (ES): m/z 584.4 (M+H). IR cm⁻¹: 2966, 1670,1634.Analytical HPLC: (4.6×150 mm Prodigy ODS (3) column eluted with 10 to100% acetonitrile in water containing 0.02% formic acid over 30minutes): 98.325% (at 8.182 minutes).

EXAMPLE 8N,β,β,3,5-Pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-(4-methylpiperazin-1-yl)-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure Method A,N,β,β,3,5-pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(90 mg, 0.179 mmol) is treated with hydroxybenzotriazole (50 mg, 0.370mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(70 mg, 0.365 mmol) in anhydrous acetonitrile (8 ml) at room temperaturefor 3 hours, followed by addition of 1-methylpiperazine (0.035 ml, 0.32mmol). After stirring at room temperature for 15 hours, the product ispurified by preparative HPLC, to provide the trifluoroacetic acid saltof the title compound as a white solid. MS (ES): m/z 584.4 (M+H). IRcm⁻¹: 2966,1671,1634 Analytical HPLC: (4.6×150 mm YMC Pack Pro C18column eluted with 10 to 100% acetonitrile in water containing 0.02%formic acid over 30 minutes): 96.63% (at 8.049 minutes) ofN,β,β,3-tetramethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-(4-methylpiperazin-1-yl)-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide,and 1.39% (at 9.352 minutes) of the other diastereomer.

EXAMPLE 91-{(2E,4S)-2,5-Dimethyl-4-[methyl(N,β,β,3-tetramethyl-L-phenylalanyl-3-methyl-L-valyl)amino]hex-2-enoyl}-D-prolyl-N-benzyl-D-prolinamide trifluoroacetic acidsalt

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3-tetramethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(100 mg, 0.205 mmol) is treated with hydroxybenzotriazole (36 mg, 0.27mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(59 mg, 0.31 mmol) in anhydrous acetonitrile (10 ml) at room temperaturefor 40 hours, followed by addition of diisopropylethylamine (0.143 ml,0.82 mmol) and a solution of L-prolyl-N-benzyl-L-prolinamidetrifluoroacetic acid salt (255 mg, 0.615 mmol) in anhydrous acetonitrile(3 ml). After stirring at room temperature for 4 hours, the product ispurified by preparative HPLC, to provide the trifluoroacetic acid saltof the title compound (126.6 mg, 69%) as a white amorphous solid. MS(ES): m/z 771.5 (M+H). IR cm⁻¹: 3306.94, 2969.17, 1673.9, 1642.27,1203.38. Analytical HPLC: (4.6×150 mm YMC Pack Pro C18 column elutedwith 10 to 100% acetonitrile in water containing 0.02% formic acid over35 minutes): 92.56% (at 17.7 minutes) of1-{(2E,4S)-2,5-dimethyl-4-[methyl(N,β,β,3-tetramethyl-L-phenylalanyl-3-methyl-L-valyl)amino]hex-2-enoyl}-D-prolyl-N-benzyl-D-prolinamide,and 6.92% (at 18.5 minutes) of the other diastereomer.

EXAMPLE 101-{(2E,4S)-2,5-Dimethyl-4-[methyl(N,β,β,3,4-pentamethyl-L-phenylalanyl-3-methyl-L-valyl)amino]hex-2-enoyl}-L-prolyl-N-benzyl-L-prolinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3,4-pentamethyl-L-phenylalanyl-N¹[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(128 mg, 0.26 mmol) is treated with hydroxybenzotriazole (42 mg, 0.32mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(70 mg, 0.36 mmol) in anhydrous acetonitrile (5 ml) at room temperaturefor 15 hours, followed by addition of diisopropylethylamine (0.068 ml,0.39 mmol) and a solution of L-prolyl-N-benzyl-L-prolinamidetrifluoroacetic acid salt (108 mg, 0.26 mmol) in anhydrous acetonitrile(5 ml). After stirring at room temperature for 24 hours, the product ispurified by preparative HPLC, to provide the trifluoroacetic acid saltof the title compound 126.6 mg, 62.0%) as a white amorphous solid. MS(ES): m/z 785.5 (M+H). IR cm⁻¹: 2967, 1634. Analytical HPLC: (4.6×150 mmYMC Pack Pro C18 column eluted with 10 to 100% acetonitrile in watercontaining 0.02% formic acid over 30 minutes): 96.35% (at 18.713minutes) of1-{(2E,4S)-2,5-dimethyl-4-[methyl(N,β,β,3,4-pentamethyl-L-phenylalanyl-3-methyl-L-valyl)amino]hex-2-enoyl}-L-prolyl-N-benzyl-L-prolinamide,and 0.73% (at 19.277 minutes) of the other diastereomer.

EXAMPLE 111-{(2E,4S)-2,5-Dimethyl-4-[methyl(N,β,β,3,5-pentamethyl-L-phenylalanyl-3-methyl-L-valyl)amino]hex-2-enoyl}-L-prolyl-N-benzyl-L-prolinamide

By using an analogous procedure described in General Procedure Method A,N,β,β,3,5-pentamethyl-L-phenylalanyl-N¹[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(83 mg, 0.166 mmol) is treated with hydroxybenzotriazole (27 mg, 0.20mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(45 mg, 0.23 mmol) in anhydrous acetonitrile (5 ml) at room temperaturefor 15 hours, followed by addition of diisopropylethylamine (0.045 ml,0.25 mmol) and a solution of L-prolyl-N-benzyl-L-prolinamidetrifluoroacetic acid salt (69 mg, 0.166 mmol) in anhydrous acetonitrile(5 ml). After stirring at room temperature for 24 hours, the product ispurified by preparative HPLC, to provide the trifluoroacetic acid saltof the title compound as a white amorphous solid. MS (ES): m/z 785.2(M+H). IR cm⁻¹: 2968,1635. Analytical HPLC: (4.6×150 mm YMC Pack Pro C18column eluted with 10 to 100% acetonitrile in water containing 0.02%formic acid over 30 minutes): 83.99% (at 13.513 minutes) of1-{(2E,4S)-2,5-dimethyl-4-[methyl(N,β,β,3,5-pentamethyl-L-phenylalanyl-3-methyl-L-valyl)amino]hex-2-enoyl}-L-prolyl-N-benzyl-L-prolinamide,and 3.02% (at 13.795 minutes) of the other diastereomer.

EXAMPLE 12N,β,β,3,4-Pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3,4-pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(100 mg, 0.199 mmol) is treated with hydroxybenzotriazole (35 mg, 0.259mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(57 mg, 0.299 mmol) in acetonitrile (10 ml) at room temperature for 15hours, followed by addition of diisopropylethylamine (0.139 ml, 0.797mmol) and 1-benzylpiperazine (0.069 ml, 0.399 mmol). After stirring atroom temperature for 6.5 hours, the product is purified by preparativeHPLC, to provide the trifluoroacetic acid salt of the title compound(136 mg, 88%) as a white solid. MS (ES): m/z 660.5 (M+H), 330.9 (2M+H).IR cm⁻¹: 3426.46, 2969.86, 1673.74, 1634.74, 1202.94. Analytical HPLC:(4.6×150 mm YMC Pack Pro C18 column eluted with 10 to 95% acetonitrilein water containing 0.02% formic acid over 25 minutes): 93.90% (at 8.3minutes).

EXAMPLE 13N,β,β,3,4-Pentamethyl-D-phenylalanyl-N¹-[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3,4-pentamethyl-D-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(100 mg, 0.199 mmol) is treated with hydroxybenzotriazole (35 mg, 0.259mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(57 mg, 0.299 mmol) in acetonitrile (10 ml) at room temperature for 15hours, followed by addition of diisopropylethylamine (0.139 ml, 0.797mmol) and 1-benzylpiperazine (0.069 ml, 0.399 mmol). After stirring atroom temperature for 6.5 hours, the product is purified by preparativeHPLC, to provide the trifluoroacetic acid salt of the title compound(113 mg, 86%) as a white solid. MS (ES): m/z 660.5 (M+H), 330.7 (M+2H).IR cm⁻¹: 3328.24, 2969.50, 1674.73, 1633.08, 1202.46, 1134.70.Analytical HPLC: (4.6×150 mm Prodigy ODS (3) column eluted with 10 to100% acetonitrile in water containing 0.02% formic acid over 25minutes): 5.04% (at 8.2 minutes), 79.40% (at 8.7 minutes).

EXAMPLE 14N,β,β,3,5-Pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3,5-pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(100 mg, 0.199 mmol) is treated with hydroxybenzotriazole (35 mg, 0.259mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(57 mg, 0.299 mmol) in acetonitrile (10 ml) at room temperature for 15hours, followed by addition of diisopropylethylamine (0.139 ml, 0.799mmol) and 1-benzylpiperazine (0.069 ml, 0.399 mmol). After stirring atroom temperature for 15 hours, the product is purified by preparativeHPLC, to provide the trifluoroacetic acid salt of the title compound(125 mg, 81%) as a white solid. MS (ES): m/z 660.6 (M+H), 331.0 (2M+H).IR cm⁻¹: 3434.21, 2968.89, 1674.58, 1632.61, 1203.08. Analytical HPLC:(4.6×150 mm YMC Pack Pro C18 column eluted with 10 to 95% acetonitrilein water containing 0.02% formic acid over 25 minutes): 93.38% at 8.6minutes

EXAMPLE 15N,β,β,β-Tetramethyl-L-phenylalanyl-N¹[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3-tetramethyl-L-phenylalanyl-N¹[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(150 mg, 0.31 mmol) is treated with hydroxybenzotriazole (54 mg, 0.4mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(59 mg, 0.46 mmol) in acetonitrile (10 ml) at room temperature for 15hours, followed by addition of diisopropylethylamine (0.143 ml, 0.82mmol) and 1-benzylpiperazine (0.043 ml,0.25 mmol). After stirring atroom temperature for 3 days, the product is purified by preparativeHPLC, to provide the trifluoroacetic acid salt of the title compound (81mg, 34%) as a yellow solid. MS (ES): m/z 646.4 (M+H), 323.8 (M+2H). IRcm⁻¹: 3428.51, 2969.56, 1674.67, 1202.94. Analytical HPLC: (4.6×150 mmYMC Pack Pro C18 column eluted with 10 to 95% acetonitrile in watercontaining 0.02% formic acid over 25 minutes): 90.90% at 6.5 minutes.

EXAMPLE 163-Chloro-N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,3-chloro-N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(121 mg, 0.238 mmol) is treated with hydroxybenzotriazole (42 mg, 0.31mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(68 mg, 0.36 mmol) in acetonitrile (10 ml) at room temperature for 15hours, followed by addition of diisopropylethylamine (0.166 ml, 0.95mmol) and 1-benzylpiperazine (0.05 ml, 0.29 mmol). After stirring atroom temperature for 24 hours, the product is purified by preparativeHPLC, to provide the trifluoroacetic acid salt of the title compound(157 mg, 85%) as a tan solid. MS (ES): m/z 666.5 (M+H), 333.8 (M+2H). IRcm⁻¹: 2970.22, 1674.58, 1637.35, 1420.27, 1203.00. Analytical HPLC:(4.6×150 mm YMC Pack Pro C18 column eluted with 10 to 95% acetonitrilein water containing 0.02% formic acid over 25 minutes): 86.28% at 7.9minutes, 10.39% at 8.1 minutes, 1.01% at 8.4 minutes.

EXAMPLE 17N,β,β,3,4-Pentamethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3,4-pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(150 mg, 0.299 mmol) is treated with hydroxybenzotriazole (53 mg, 0.384mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(86 mg, 0.448 mmol) in acetonitrile (10 ml) at room temperature for 15hours, followed by addition of diisopropylethylamine (0.208 ml, 1.2mmol) and L-proline methyl ester hydrochloride (59 mg, 0.36 mmol). Afterstirring at room temperature for 4 hours, the product is purified bypreparative HPLC, to provide the trifluoroacetic acid salt of the titlecompound (208 mg, 96%) as a white solid. MS (ES): m/z 613.4 (M+H). IRcm⁻¹: 3395.09, 2968.00, 1747.41, 1674.85, 1630.23, 1202.70. AnalyticalHPLC: (4.6×150 mm YMC Pack Pro C18 column eluted with 10 to 95%acetonitrile in water containing 0.02% formic acid over 25 minutes):87.55% at 11.2 minutes.

EXAMPLE 18N,β,β,3,4-Pentamethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure II,methyl(2S)-1-{(2E,4S)-4-[((2S)-2-{[(2S)-3-(3,4-dimethylphenyl)-3-methyl-2-(methylamino)butanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethylhex-2-enoyl}pyrrolidine-2-carboxylatetrifluoroaceti acid salt (140 mg, 0.19 mmol, obtained from Example 17)in methanol (1.5 ml) and water (0.5 ml) is treated with lithiumhydroxide (1 mmol, aqueous solution). After stirring at room temperaturefor 3 hours, the product is purified by preparative HPLC, to provide thetrifluoroacetic acid salt of the title compound (147 mg, 100%) as awhite solid. MS (ES): m/z 599.5 (M+H). IR cm⁻¹: 3431.56, 2968.23,1729.35, 1201.76. Analytical HPLC: (4.6×150 mm YMC Pack Pro C18 columneluted with 10 to 95% acetonitrile in water containing 0.02% formic acidover 25 minutes): 94.83% at 10.2 minutes.

EXAMPLE 19N,β,β,3,5-Pentamethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,N,β,β,3,5-pentamethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(150 mg, 0.299 mmol) is treated with hydroxybenzotriazole (53 mg, 0.389mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(86 mg, 0.448 mmol) in acetonitrile (10 ml) at room temperature for 15hours, followed by addition of diisopropylethylamine (0.208 ml, 1.2mmol) and L-proline methyl ester hydrochloride (59 mg, 0.36 mmol). Afterstirring at room temperature for 15 hours, the product is purified bypreparative HPLC, to provide the trifluoroacetic acid salt of the titlecompound (106 mg, 49%) as a white solid. MS (ES): m/z 613.4 (M+H). IRcm⁻¹: 3388.60, 2967.14, 1747.20, 1676.16, 1631.95, 1202.88. AnalyticalHPLC: (4.6×150 mm YMC Pack Pro C18 column eluted with 10 to 95%acetonitrile in water containing 0.02% formic acid over 25 minutes):81.54% at 11.4 minutes.

EXAMPLE 20N,β,β,3,5-Pentamethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure II,methyl(2S)-1-{(2E,4S)-4-[((2S)-2-{[(2S)-3-(3,5-dimethylphenyl)-3-methyl-2-(methylamino)butanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethylhex-2-enoyl}pyrrolidine-2-carboxylatetrifluoroacetic acid salt (58 mg, 0.095 mmol obtained from Example 19)in methanol (1.5 ml) and water (0.5 ml) is treated with lithiumhydroxide (1 mmol, aqueous solution). After stirring at room temperaturefor 3 hours, the product is purified by preparative HPLC, to provide thetrifluoroacetic acid salt of the title compound (35 mg, 52%) as a whitesolid. MS (ES): m/z 599.4 (M+H). Analytical HPLC: (4.6×150 mm YMC PackPro C18 column eluted with 10 to 95% acetonitrile in water containing0.02% formic acid over 25 minutes): 94.37% at 10.3 minutes.

EXAMPLE 21N,β,β,3-Tetramethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA, N,β,β,3-tetramethyl-L-phenylalanyl-N¹—[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(100 mg, 0.205 mmol) is treated with hydroxybenzotriazole (36 mg, 0.27mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(59 mg, 0.31 mmol) in acetonitrile (10 ml) at room temperature for 15hours, followed by addition of diisopropylethylamine (0.215 ml, 1.23mmol) and L-proline methyl ester hydrochloride (61 mg, 0.37 mmol). Afterstirring at room temperature for 2 days, the product is purified bypreparative HPLC, to provide the trifluoroacetic acid salt of the titlecompound (75 mg, 51%) as a yellow solid. MS (ES): m/z 599.4 (M+H). IRcm⁻¹: 2969.22, 1748.56, 1676.30, 1628.91, 1204.24. Analytical HPLC:(4.6×150 mm YMC Pack Pro C18 column eluted with 10 to 95% acetonitrilein water containing 0.02% formic acid over 25 minutes): 85.72% at 9.8minutes, 1.23% at 10.8 minutes.

EXAMPLE 22N,β,β,3-Tetramethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure II,methyl(2S)-1-{(2E,4S)-4-[((2S)-3,3-dimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3(3-methylphenyl)butanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethylhex-2-enoyl}pyrrolidine-2-carboxylatetrifluoroacetic acid salt (141 mg, 0.198 mmol, obtained from Example 21)in methanol (1.5 ml) and water (0.5 ml) is treated with lithiumhydroxide (1 mmol, aqueous solution). After stirring at room temperaturefor 15 hours, the product is purified by preparative HPLC, to providethe trifluoroacetic acid salt of the title compound (106 mg, 77%) as awhite solid. MS (ES): m/z 585.3 (M+H). IR cm⁻¹: 2969, 1677, 1629, 1451,1415, 1203. Analytical HPLC: (4.6×150 mm YMC Pack Pro C18 column elutedwith 10 to 100% acetonitrile in water containing 20 mM ammonium acetateover 27 minutes): 99.50% at 14.3 minutes.

EXAMPLE 233-Chloro-N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure I MethodA,3-chloro-N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N¹,3-dimethyl-L-valinamide(119 mg, 0.234 mmol) is treated with hydroxybenzotriazole (41 mg, 0.305mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(67 mg, 0.35 mmol) in acetonitrile (10 ml) at room temperature for 15hours, followed by addition of diisopropylethylamine (0.163 ml, 0.94mmol) and L-proline methyl ester hydrochloride (47 mg, 0.28 mmol). Afterstirring at room temperature for 15 hours, the product is purified bypreparative HPLC, to provide the trifluoroacetic acid salt of the titlecompound (134 mg, 78%) as a white solid. MS (ES): m/z 619.4 (M+H). IRcm⁻¹: 2968.43, 1677.81, 1634.03, 1416.23, 1203.94, 1175.63. AnalyticalHPLC: (4.6×150 mm Capcell C18 column eluted with 10 to 95% acetonitrilein water containing 0.02% formic acid over 25 minutes): 77.08% at 9.8minutes, 8.11% at 10.5 minutes.

EXAMPLE 243-Chloro-N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

By using an analogous procedure described in General Procedure II,methyl(2S)-1-{(2E,4S)-4-[((2S)-2-{[(2S)-3-(3-chlorophenyl)-3-methyl-2-(methylamino)butanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethylhex-2-enoyl}pyrrolidine-2-carboxylatetrifluoroacetic acid salt (84 mg, 0.135 mmol, obtained from Example 23)in methanol (1 ml) and water (0.5 ml) is treated with lithium hydroxide(0.5 mmol). After stirring at room temperature for 6.5 hours, theproduct is purified by preparative HPLC, to provide the trifluoroaceticacid salt of the title compound (45 mg, 46%) as a white solid. MS (ES):m/z 605.5 (M+H). Analytical HPLC: (4.6×150 mm YMC Pack Pro C18 columneluted with 10 to 95% acetonitrile in water containing 0.02% formic acidover 25 minutes): 95.87% at 9.9 minutes, 3.70% at 10.4 minutes.

EXAMPLE 25(E,4S)-4-[((2S)-3,3-dimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-N-hydroxy-2,5-dimethyl-2-hexenamide

As described in General Procedure I Method B, toN,β,β-trimethyl-L-phenylalanyl-N¹[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(75 mg) in acetonitrile (5.0 mL) at 25° C. is added1-hydroxybenzotriazole hydrate (26 mg) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (41 mg).After 2 hours 50% aqueous hydroxylamine (0.38 mL) is added. After 18hours the reaction mixture is concentrated in vacuo, dissolved inacetonitrile/water, and purified by reverse phase HPLC (0.01% aqueoustrifluoroacetic acid/acetonitrile gradient system) to give thetrifluoracetic acid salt of the title compound as a white powder (30mg). MS (ES): m/z 489.0 (M+H).

EXAMPLE 26N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-(4-methyl-1-piperazinyl)-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamide

As described in General Procedure I Method B toN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(50 mg) in acetonitrile (5.0 mL) at 25° C. is added1-hydroxybenzotriazole hydrate (17 mg) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (27 mg).After 2 hours N-methylpiperazine (0.047 mL, 0.42 mmol) is added. After18 hours the reaction mixture is concentrated in vacuo, dissolved inacetonitrile/water, and purified by reverse phase HPLC (0.01% aqueoustrifluoroacetic acid/acetonitrile gradient system) to give thetrifluoracetic acid salt of the title compound as a white powder (58mg). MS (ES): m/z 556.42137 (M+H). (calc'd exact mass=555.41507)

EXAMPLE 27(2S)-N-[(1S,2E)-1-isopropyl-3-methyl-4-(4-morpholinyl)-4-oxo-2-butenyl]-N,3,3-trimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamide

As described in General Procedure I Method B, toN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(30 mg) in acetonitrile (5.0 mL) at 25° C. is added1-hydroxybenzotriazole hydrate (10 mg) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (16 mg).After 2 hours morpholine (0.022 mL, 0.25 mmol) is added. After 18 hoursthe reaction mixture is concentrated in vacuo, dissolved inacetonitrile/water, and purified by reverse phase HPLC (0.01% aqueoustrifluoroacetic acid/acetonitrile gradient system) to give thetrifluoracetic acid salt of the title compound as a white powder (37mg). MS (ES): m/z 543.6 (M+H).

EXAMPLE 281-{(2E,4S)-2,5-dimethyl-4-[methyl(N-methyl-3-phenyl-L-valyl-3-methyl-L-valyl)amino]hex-2-enoyl}-D-prolyl-N-benzyl-D-prolinamide

As described in General Procedure I Method B, toN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(250 mg) in acetonitrile (25 mL) at 25° C. is added1-hydroxybenzotriazole hydrate (85 mg) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (135 mg).After 21 hours a solution of D-prolyl-N-benzyl-D-prolinamidetrifluoroacetic acid (660 mg, 1.59 mmol) and diisopropylethylamine (0.37mL) in acetonitrile (3 mL) is added. After 18 hours the reaction mixtureis concentrated in vacuo, dissolved in acetonitrile/water, and purifiedby reverse phase HPLC (water/acetonitrile gradient system) to give thetitle compound as a white powder. MS (ES): m/z 757.50073 (M+H). (calc'dexact mass=756.49409)

REFERENCE EXAMPLE 1 N-methoxy-N-methyl-2-phenylacetamide

Based on a procedure found in the literature (Guanti, G.; Banfi, L.;Riva, R. Tetrahedron 50(41), 1994, 11945-11966), a solution ofphenylacetic acid (10 g, 73 mmol) in tetrahydrofuran (150 mL) iscombined with a solution of N,O-dimethylhydroxylamine hydrochloride (13g, 130 mmol) in water (150 mL). The pH is adjusted to 4.5 by theaddition of 1 N aqueous sodium hydroxide solution. To the mixture isadded 1-(3-dimethylaminopropyl)-3-ethylcarbodiimine hydrochloride (36 g,190 mmol) as a solution in water (510 mL) dropwise over 30 minutes. Overthe course of the addition, pH is maintained at 4.5 by the addition of 1N aqueous sodium hydroxide solution. The reaction mixture is allowed tostir for 3 hours at room temperature and is then saturated with sodiumchloride and extracted thrice with ethyl acetate. The combined extractsare washed with saturated aqueous sodium hydrogen carbonate (×2), water,and saturated aqueous sodium chloride solution, dried over anhydroussodium sulfate, and concentrated under reduced pressure to yield agolden liquid (13 g, 100%). MS (ES⁺): (M+H)=180.1

REFERENCE EXAMPLE 2 2-phenyl-1-(1,3-thiazol-2-yl)ethanone

Following the method of Itako et al. (Irako, N.; Hamada, Y.; Shioiri, T.Tetrahedron 48(35), 1992, 7251-7264), a solution ofN,N,N′,N′-tetramethylenediamine (9.2 mL, 61 mmol) in tetrahydrofuran(120 mL) is cooled to −78° C. (dry ice/acetone bath) while stirringunder a nitrogen atmosphere. n-butyllithium (1.6 M in hexanes, 38 mL, 61mmol) is added via syringe, followed by 2-bromothiazole (5.5 mL, 61mmol), which is also added via syringe. The reaction mixture is stirredfor 2 hours at −78° C., then N-methoxy-N-methyl-2-phenylacetamide (9.1g, 51 mmol, from Reference Example 1) is added as a solution intetrahydrofuran (120 mL). The resulting reaction mixture is stirred for30 minutes at −78° C., then at −10° C. (NaCl/ice bath) for 2 hoursbefore being quenched by the addition of 1 M potassium bisulfatesolution (300 mL). The mixture is extracted thrice with diethyl ether.The combined extracts are washed with water and saturated aqueous sodiumchloride solution, dried over anhydrous magnesium sulfate andconcentrated under reduced pressure to a brown tar. The crude materialis purified by flash chromatography (silica gel, hexanes/ethyl acetate)to give an orange-rust colored liquid (3.5 g, 35%). MS (ES⁺):(M+H)=204.0

REFERENCE EXAMPLE 3 (1R)-2-phenyl-1-(1,3-thiazol-2-yl)ethanol

Following the method of Itako et al. (Irako, N.; Hamada, Y.; Shioiri, T.Tetrahedron 48(35), 1992, 7251-7264), to a 0° C. (ice-water bath)solution of (+)-B-chlorodiisopinocamphenylborane (+-DIP-Chloride, 3.9 g,12 mmol) in diethyl ether (3 mL) is added dropwise a solution of2-phenyl-1-(1,3-thiazol-2-yl)ethanone (0.83 g, 4.1 mmol, from ReferenceExample 2) in diethyl ether (17 mL). The mixture is stirred for 6 hoursat 0° C. and then is allowed to sit in a −5° C. freezer overnight. Thenext day, 10% aqueous sodium hydroxide solution (10 mL) is added at 0°C. with stirring. Aqueous hydrogen peroxide solution (30%, 1.5 mL) isthen added and the mixture is allowed to stir at room temperature for 2hours. The biphasic mixture is diluted with water (50 mL). The twolayers are partitioned and the aqueous phase is saturated with potassiumcarbonate and then extracted thrice with diethyl ether. The combinedextracts are washed with saturated aqueous sodium chloride solution,dried over anhydrous sodium sulfate, and concentrated under reducedpressure to give a bright golden oil, which is purified by flashchromatography (silica gel, hexanes/ethyl acetate). A light yellow solid(0.60 g, 72%) is provided. MS (ES⁺): (M+H)=206.0

REFERENCE EXAMPLE 4 (1S)-2-phenyl-1-(1,3-thiazol-2-yl)ethylamine

Following the method of Itako et al. (Irako, N.; Hamada, Y.; Shioiri, T.Tetrahedron 48(35), 1992, 7251-7264), a solution of(1R)-2-phenyl-1-(1,3-thiazol-2-yl)ethanol (1.3 g, 6.3 mmol, fromReference Example 3) in tetrahydrofuran (60 mL) is cooled to 0° C.(ice-water bath). To the solution is added successivelytriphenylphosphine (1.8 g, 6.9 mmol), diethylazodicarboxylate (1.1 mL,6.9 mmol), and diphenylphosphoryl azide (DPPA, 1.6 mL, 6.9 mmol).Following the addition of reagents, the reaction mixture is allowed towarm to room temperature and stir for 47 hours. Following this period,the mixture is concentrated under reduced pressure and purified by flashchromatography (silica gel, hexanes/toluene/diethyl ether) to give 1.2 gof (1S)-2-phenyl-1-(1,3-thiazol-2-yl)ethylazide as a pale blond oilcontaminated with DPPA (detected by ¹H NMR).

To a 55° C. (oil bath) solution of triphenylphosphine (1.6 g, 6.3 mmol)in tetrahydrofuran (10 mL) is added(1S)-2-phenyl-1-(1,3-thiazol-2-yl)ethylazide (contaminated with DPPA,1.2 g, 5.2 mmol) as a solution in tetrahydrofuran (10 mL). The reactionmixture is allowed to stir for 2 hours at this temperature when aqueousammonium hydroxide (14 mL) is added. Stirring at 55° C. is continued for3 hours before the reaction mixture is stored in a −5° C. freezerovernight. The next morning, the reaction mixture is diluted with water(80 mL) and extracted twice with diethyl ether. The combined extractsare washed with 10% aqueous hydrochloric acid (70 mL). The acidicaqueous phase is cooled in an ice-water bath and then basified with 10%aqueous sodium hydroxide to pH 12. The basic phase is then extractedthrice with dichloromethane. The combined extracts are dried overanhydrous magnesium sulfate, filtered, and concentrated under reducedpressure to provide a 0.67 g (52% over 2 steps) of a slightly blond oil.MS (ES⁺): (M+H)=205.0

REFERENCE EXAMPLE 5 Tert-butyl(2S)-2-{(1R,2R)-1-hydroxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidine-1-carboxylate

In following the procedure of Shioiri et al. (Shioiri, T.; Hayashi, K.;Hamada, Y. Tetrahedron. 49(9), 1993, 1913-1924), to a 0° C. (ice-waterbath) solution of (4R,5S)-4-methyl-5-phenyl-3-propionyl-2-oxazolidinone(6.4 g, 27 mmol, Fluka) in dichloromethane (110 mL) is addedtriethylamine (4.3 mL, 31 mmol). Di-n-butylboron triflate (1.0 Msolution in dichloromethane, 29 mL, Aldrich) is added dropwise and theresulting mixture is allowed to stir for 45 minutes at 0° C., thencooled to −78° C. (dry ice/acetone bath).N-(tert-butoxycarbonyl)-L-prolinal (3.1 g, 16 mmol, Omega) is addeddropwise as a solution in dichloromethane (16 mL) and the reactionmixture is allowed to stir for 2 hours at −78° C. The reaction vessel isthen stored in a 0° C. freezer overnight. The next morning, the reactionmixture is warmed to room temperature over 15 minutes and then quenchedby the addition of pH 7 phosphate buffer (55 mL) followed by methanol(80 mL). The mixture is then cooled to 0° C. (ice-water bath) and 30%hydrogen peroxide-methanol (1:2, 96 mL) is added dropwise to keep theinternal temperature less than 10° C. After stirring for 1 hour in anice-water bath, to the reaction mixture is added water (55 mL).Volatiles are removed under reduced pressure and the aqueous residue isextracted thrice with diethyl ether. The combined extracts are washedwith 1 M potassium bisulfate solution, water, saturated aqueous sodiumbicarbonate solution, and saturated aqueous sodium chloride solution,dried over anhydrous sodium sulfate, and concentrated under reducedpressure. The crude viscous pale yellow oil (9.5 g) is purified by flashchromatography (hexanes/ethyl acetate) to give 4.6 g (67%) of a whitefoam. MS (ES⁺): (M+H)=433.3

EXAMPLE 29N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[(2S)-2-((1R,2R)-1-methoxy-2-methyl-3-oxo-3-{[(1S)-2-phenyl-1-(1,3-thiazolyl-2-yl)ethyl]amino}propyl)pyrrolidin-1-yl]-3-methyl-4-oxobut-2-dimethyl-L-valinamide

In following the procedure of Shioiri et al. (Shioiri, T.; Hayashi, K.;Hamada, Y. Tetrahedron 49(9), 1993,1913-1924), to a 0° C. (ice-waterbath) solution of tert-butyl(2S)-2-{(1R,2R)-1-hydroxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidine-1-carboxylate(0.89 g, 2.1 mmol, from Reference Example 5) is added dropwise aqueoushydrogen peroxide solution (30%, 1.1 mL, 11 mmol), followed by 0.4 Maqueous lithium hydroxide solution (11 mL, 4.2 mmol). After 2½ hours ofstirring at 0° C., 1 M aqueous sodium sulfite solution (12 mL) is addeddropwise, and the quenched reaction mixture is stirred over the weekendat room temperature. The mixture is poured into ice-cold saturatedaqueous sodium bicarbonate solution (80 mL) and extracted thrice withdichloromethane to remove the chiral auxiliary. The basic aqueous phaseis cooled to 0° C., acidified to pH 2 with 1 M aqueous potassiumbisulfate solution, saturated with sodium chloride, and extracted thricewith diethyl ether. The combined extracts are washed with water andsaturated aqueous sodium chloride solution, dried over anhydrous sodiumsulfite, and concentrated under reduced pressure to give(2R,3R,2′S)-3-(N-tert-butoxycarbonyl-2′-pyrrolidinyl)-3-hydroxy-2-methylpropanoicacid as a flocculent white solid (0.56 g, 98%).

In following the procedure of Shioiri et al. (Shioiri, T.; Hayashi, K.;Hamada, Y. Tetrahedron. 49(9), 1993, 1913-1924), to a solution of(2R,3R,2′S)-3-(N-tert-butoxycarbonyl-2′-pyrrolidinyl)-3-hydroxy-2-methylpropanoicacid (0.56 g, 2.0 mmol) in tetrahydrofuran (10 mL) is added iodomethane(1.9 mL, 30 mmol). The mixture is cooled to 0° C. (ice-water bath) whilestirring under a nitrogen atmosphere. Sodium hydride (60% dispersion inmineral oil, 0.32 g, 8.0 mmol) is added in portions to keep the reactionmixture from frothing excessively. Stirring is continued for 2 hours at0° C. and then the reaction vessel is stored for 3 days in a −5° C.freezer. The mixture is quenched by the addition of saturated aqueoussodium bicarbonate solution (20 mL) and then extracted with ether toremove impurities and mineral oil. The basic aqueous phase is acidifiedto pH 2 with 1 M potassium bisulfate solution and then extracted twicewith ethyl acetate. The combined extracts are washed with 5% aqueoussodium thiosulfite solution and saturated aqueous sodium chloridesolution, dried over anhydrous sodium sulfite, and concentrated underreduced pressure to give(2R,3R,2′S)-3-(N-tert-butoxycarbonyl-2′-pyrrolidinyl)-3-methoxy-2-methylpropanoicacid (Boc-(2R,3R,4S)-dolaproine) as a clear, colorless semisolid (0.41g, 72%).

In following the procedure of Shioiri et al. (Shioiri, T.; Hayashi, K.;Hamada, Y. Tetrahedron. 49(9), 1993, 1913-1924), a solution of(1S)-2-phenyl-1-(1,3-thiazol-2-yl)ethylamine (0.20 g, 1.0 mmol, fromReference Example 4) and(2R,3R,2′S)-3-(N-tert-butoxycarbonyl-2′-pyrrolidinyl)-3-methoxy-2-methylpropanoicacid (0.28 g, 1.0 mmol) in anhydrous dimethylformamide (3 mL) is cooledto 0° C. (ice-water bath) while stirring under a nitrogen atmosphere.Diethyl cyanophosphonate (0.16 mL, 1.05 mmol) is added dropwise viasyringe, followed by triethylamine (0.14 mL, 1.0 mmol). Stirring iscontinued for two hours at room temperature and then at room temperaturefor three days. The reaction mixture is diluted with 30 mL of ethylacetate/toluene (2:1) and washed with 1 M aqueous potassium bisulfatesolution, water, saturated aqueous sodium bicarbonate solution, andsaturated aqueous sodium chloride solution. The organic phase is driedover anhydrous sodium sulfate, decanted, and concentrated under reducedpressure to give 0.47 g (100%) ofBoc-(2R,3R,4S)-dolaproine-(S)-dolaphenine as a light yellow foam.

To a solution of crude Boc-(2R,3R,4S)-dolaproine-(S)-dolaphenine (0.23g, 0.49 mmol) in dichloromethane (1.5 mL) is added trifluoroacetic acid(0.45 mL). The reaction mixture is allowed to stir at room temperatureover the weekend. The solvent and excess trifluoroacetic acid areremoved under reduced pressure. The crude product is combined with thatfrom an identical run and purified by reverse-phase HPLC (Prodigy ODS3column, acetonitrile/water) to give 0.18 g (50% for 2 runs) of(2R,3R,4S)-dolaproine-(S)-dolaphenine as a white solid.

A mixture ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(94 mg, 0.20 mmol, WO 99/32509) and(2R,3R,4S)-dolaproine-(S)-dolaphenine (66 mg, 0.18 mmol) in anhydrousdimethylformamide (1 mL) are reacted with diethyl cyanophosphonate (33μL, 0.22 mmol) in the presence of triethylamine (31 μL, 0.22 mmol).After stirring for 2 days, the reaction mixture is quenched with a fewdrops of water and immediately purified by reverse-phase HPLC (ProdigyODS3 column, acetonitrile/water/trifluoroacetic acid) to give the titlecompound as a TFA salt (84 mg). MS (ES⁺): (M+H) 829.6

EXAMPLE 30N,β,β-trimethyl-L-phenylalanyl-N¹-((1S,2E)-4-{(2S)-2-[(1R,2R)-1,3-dimethoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl}-1-isopropyl-3-methyl-4-oxobut-2-enyl)-N¹,3-dimethyl-L-valinamide

To a 0° C. (ice-water bath) solution of(2R,3R,2′S)-3-(N-tert-butoxycarbonyl-2′-pyrrolidinyl)-3-methoxy-2-methylpropanoicacid (0.20 g, 0.70 mmol, from Example 29) in methanol (2 mL) and diethylether (1 mL) is added trimethylsilyl diazomethane (2.0 M in hexanes,approx. 0.5-0.6 mL, Aldrich) until the yellow color of the reagentpersisted. Then the reaction mixture is allowed to stir at roomtemperature for 10 minutes, after which the volatiles are removed underreduced pressure to a give 0.21 g (>100%) of crude methyl(2R,3R,2′S)-3-(N-tert-butoxycarbonyl-2′-pyrrolidinyl)-3-methoxy-2-methylpropanoate.

Without further purification, crude methyl(2R,3R,2′S)-3-(N-tert-butoxycarbonyl-2′-pyrrolidinyl)-3-methoxy-2-methylpropanoate(0.21 g, 0.70 mmol max.) is taken up in dioxane (1 mL). To the solution,hydrochloric acid (4N in dioxanes, 2 mL, Aldrich) is added, and theresulting mixture is allowed to stir overnight under a nitrogenatmosphere. The next morning, the dioxane solvent is evaporated underreduced pressure to leave methyl(2R,3R,2′S)-3-(2′-pyrrolidinyl)-3-methoxy-2-methylpropanoatehydrochloride as a straw-colored oil (0.21 g,>100%), which is used inthe next step without further purification.

Using General Procedure Method A, to a solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(0.40 g, 0.84 mmol, WO 99/32509) and methyl(2R,3R,2′S)-3-(2′-pyrrolidinyl)-3-methoxy-2-methylpropanoatehydrochloride (0.70 mmol max) in anhydrous dimethylformamide (7 mL), isadded hydroxybenzotriazole (0.19 g, 1.4 mmol),1-(3-dimethylaminopropyl)-3-ethylcarbodiimine hydrochloride (0.27 g, 1.4mmol), and N-methylmorpholine (0.15 mL, 1.4 mmol) under an inertatmosphere. The resulting mixture is stirred overnight at roomtemperature under an inert atmosphere. The mixture is diluted withwater, and the aqueous layer is extracted with diethyl ether (3 times).The combined extracts are washed with saturated aqueous sodiumbicarbonate solution and saturated aqueous sodium chloride. The extractsare dried over anhydrous sodium sulfate, filtered and concentrated invacuo to give 0.57 g (>100%) of crude product, of which 0.24 g ispurified on reverse-phase HPLC (Prodigy ODS3,acetonitrile/water/trifluoroacetic acid) to give 100 mg of the titlecompound as a TFA salt. TOF MS (ES⁺): (M+H)=657.2

EXAMPLE 31 N-methyl-3-phenyl-L-valyl-N¹—[(1S,2E)-1-isopropyl-4-((2S)-2-{(1R,2R)-1-methoxy-2-methyl-3-oxo-[(2-phenethyl)amino]propyl}pyrrolidin-1-yl)-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

(2R,3R,2′S)-3-(N-tert-butoxycarbonyl-2′-pyrrolidinyl)-3-methoxy-2-methylpropanoicacid (0.20 g, 0.70 mmol, from Example 29) is coupled to phenethylamine(0.18 mL, 1.4 mmol) using General Procedure I Method A to give2-(1-methoxy-2-phenethyloxycarbonyl-propyl)-pyrrolidine-1-carboxylicacid tert-butyl ester (0.29 g,>100%) as viscous light brown oil.

Without further purification,2-(1-Methoxy-2-phenethyloxycarbonyl-propyl)-pyrrolidine-1-carboxylicacid tert-butyl ester (0.29 g, 0.70 mmol maximum) is taken up in dioxane(2 mL). To the solution is added hydrochloric acid (4N in dioxanes, 3mL, 12 mmol). The reaction mixture is stirred for two days at roomtemperature and then concentrated under reduced pressure to give thedeprotected product (0.31 g, 0.70 mmol maximum) as a yellow semi-solid.

Without further purification, the crude material from the previousreaction (0.70 mmol maximum) is coupled toN,β,β-trimethyl-L-phenylalanyl-N¹[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(0.36 g, 0.77 mmol) using General Procedure I Method A. A portion of thecrude product is purified by reverse-phase HPLC (Prodigy ODS3 column,acetonitrile/water/formic acid) to give the title compound (160 mg) as aformic acid salt. MS (ES⁺): (M+H)=746.4

EXAMPLE 32N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-4-((2S)-2-{(1R,2R)-1-methoxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidin-1-yl)-3-methyl-4-oxovalinamide

Following a literature method (Diem, M. J.; Burow, D. F.; Fry, J. L.Journal of Organic Chemistry 42(10), 1977, 1801-1802), to a solution oftert-butyl(2S)-2-{(1R,2R)-1-hydroxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidine-1-carboxylate(0.70 g, 1.6 mmol, from Reference Example 5) in anhydrousdichloromethane (5 mL) is addedN,N,N′,N′-tetramethyl-1,8-naphthalenediamine (0.62 g, 2.9 mmol),followed by trimethyloxonium tetrafluoroborate (0.35 g, 2.4 mmol). Theresulting reaction mixture is stirred for 24 hours at room temperature,quenched by the addition of ice water (25 mL), and then filtered througha diatomaceous earth pad. The filtrate is evaporated under reducedpressure and then partitioned between diethyl ether and water. Theaqueous phase is extracted thrice with diethyl ether. The combinedorganic extracts are washed with 1 M aqueous potassium bisulfatesolution, water, saturated aqueous sodium bicarbonate solution, andsaturated aqueous sodium chloride solution, dried over anhydrous sodiumsulfate, and concentrated under reduced pressure to give 0.69 g (97%) oftert-butyl(2S)-2-{(1R,2R)-1-methoxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidine-1-carboxylateas a foam.

Without further purification, crude tert-butyl(2S)-2-{(1R,2R)-1-methoxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidine-1-carboxylateis dissolved in anhydrous dichloromethane (5 mL). Trifluoroacetic acid(TFA, 1 mL) is added and the resulting reaction mixture is stirred for 4hours at room temperature. Excess TFA and solvent are evaporated underreduced pressure, and the crude product is purified by reverse-phaseHPLC (Prodigy ODS3 column, acetonitrile/water) to give 0.41 g (76%) of(2S)-2-{(1R,2R)-1-methoxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidineas a hard white foam.

Using General Procedure I Method A,N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(0.23 g, 0.49 mmol) is coupled to(2S)-2-{(1R,2R)-1-methoxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidine(0.17 g, 0.49 mmol). A portion of the crude material is purified byreverse-phase HPLC (Prodigy ODS3 column,acetonitrile/water/trifluoroacetic acid) to give 93 mg of the TFA saltof the title compound as a white powder. MS (ES⁺): (M+H)=802.4

EXAMPLE 33 MethylN,β,β-trimethyl-L-phenylalanyl-3-methyl-L-valyl-N-methyl-L-valyl-L-prolinate

Using General Procedure I Method A,(2S)-(tert-Butoxycarbonyl-methyl-amino)-3-methyl-butyric acid (0.37 g,1.6 mmol) is coupled to L-proline methyl ester hydrochloride (0.26 g,1.6 mmol) to give1-[2-(tert-butoxycarbonyl-methyl-amino)-3-methyl-butyryl]-pyrrolidine-2-carboxylicacid methyl ester (0.51 g, 93%) as a white solid.

Without further purification, this material is taken up in anhydrousdichloromethane (5 mL). Trifluoroacetic acid (1.4 mL) is added to thesolution, which is then allowed to stir for 2 days at room temperature.Volatiles are evaporated under reduced pressure to give1-(3-methyl-2-methylamino-butyryl)-pyrrolidine-2-carboxylic acid methylester.

To a solution of (2S)-(tert-butoxycarbonyl-methyl-amino)-tert-leucine(0.69 g, 3.0 mmol) and the crude material from the previous step (0.54g, 1.5 mmol maximum) in anhydrous dichloromethane (7 mL) is addedbenzotriazole-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate(PyBOP, 1.2 g, 2.4 mmol) and diisopropylethylamine (0.86 mL, 5.1 mmol)under an inert atmosphere. The resulting reaction mixture is stirred atroom temperature for 3 days. The mixture is diluted ethylacetate/toluene (2:1) and washed with 1M potassium bisulfate solution,water, saturated aqueous sodium bicarbonate solution, and saturatedaqueous sodium chloride, dried over anhydrous sodium sulfate, filteredand concentrated in vacuo. The residue,1-{2-[(2-tert-butoxycarbonylamino-3,3-dimethyl-butyryl)-methyl-amino]-3-methyl-butyryl}-pyrrolidine-2-carboxylicacid methyl ester, a dark straw colored oil, is submitted to the nextstep without further purification.

The crude material (1.5 mmol maximum) from the previous step is taken upin anhydrous dichloromethane (10 mL). Trifluoroacetic acid (2 mL) isadded to the solution, which is then allowed to stir for 2.5 hours atroom temperature. Excess trifluoroacetic acid and dichloromethane areevaporated under reduced pressure and the residue is purified byreverse-phase HPLC (Prodigy ODS3 column, acetonitrile/water) to give1-{2-[(2-amino-3,3-dimethyl-butyryl)-methyl-amino]-3-methyl-butyryl}-pyrrolidine-2-carboxylicacid methyl ester (0.50 g, 94% over three steps) as a hard white foam.

Using General Procedure I Method A, the product from the previousreaction (0.50 g, 1.4 mmol) is coupled to(2S)-2-[(tert-butoxycarbonyl)(methyl)amino]-3-methyl-3-phenylbutanoicacid (0.43 g, 1.4 mmol) to give1-[2-({2-[2-(tert-butoxycarbonyl-methyl-amino)-3-methyl-3-phenyl-butyrylamino]-3,3-dimethyl-butyryl}-methyl-amino)-3-methyl-butyryl]-pyrrolidine-2-carboxylicacid methyl ester. The crude coupling product is submitted to thefollowing step without further purification. The material (0.7 mmolmaximum) from the previous step is taken up in anhydrous dichloromethane(5 mL). Trifluoroacetic acid (1.5 mL) is added to the solution, which isthen allowed to stir for 4 hours at room temperature. Excesstrifluoroacetic acid and dichloromethane are evaporated under reducedpressure and the residue is purified by reverse-phase HPLC (Prodigy ODS3column, acetonitrile/water/trifluoroacetic acid) to give 0.24 g of theTFA salt of the title compound as a hard white foam. MS (ES⁺):(M+H)=545.4

EXAMPLE 34N,β,β-trimethyl-L-phenylalanyl-3-methyl-L-valyl-N-methyl-L-valyl-L-prolyl-N-benzyl-L-prolinamide

To a solution of methylN,β,β-trimethyl-L-phenylalanyl-3-methyl-L-valyl-N-methyl-L-valyl-L-prolinate(0.7 mmol maximum, from Example 33) in tetrahydrofuran (2 mL), methanol(2 mL), and water (1 mL) is added lithium hydroxide monohydrate (50 mg).The reaction mixture is stirred in a 45° C. oil bath overnight. On thefollowing day, the reaction mixture is allowed to cool to roomtemperature. Following concentration under reduced pressure, the crudeproduct is purified by reverse-phase HPLC (Prodigy ODS3 column,acetonitrile/water/trifluoroacetic acid) to give 0.22 mg of thetrifluoroacetic acid salt ofN,β,β-trimethyl-L-phenylalanyl-3-methyl-L-valyl-N-methyl-L-valyl-L-prolineas a white powder.

Using General Procedure I Method A, N-(tert-butoxycarbonyl)-L-proline(1.0 g, 4.6 mmol) is coupled to benzylamine (0.55 g, 5.1 mmol) to give1.2 g (86%) of 2-benzylcarbamoyl-pyrrolidine-1-carboxylic acidtert-butyl ester, which is submitted to the following step withoutfurther purification.

A sample of the material from the previous step (0.43 g, 1.4 mmol) isdissolved in dioxane (2 mL), and to the solution is added hydrochloricacid (4N in dioxanes, 3 mL, 12 mmol). The reaction mixture is allowed tostir for 2 hours at room temperature and then concentrated under reducedpressure to provide a viscous blond oil. Ether is added and evaporatedsequentially three times to give N-benzyl-L-prolinamide hydrochloride asa pale yellow paste.

Using General Procedure I Method A, N-benzyl-L-prolinamide hydrochloride(0.18 g, 0.76 mmol) is coupled to the trifluoroacetic acid salt ofN,β,β-trimethyl-L-phenylalanyl-3-methyl-L-valyl-N-methyl-L-valyl-L-proline(0.11 g, 0.15 mmol, WO 99/32509). After stirring overnight at roomtemperature, the reaction mixture is purified without an aqueous workupby reverse-phase HPLC (acetonitrile/water/trifluoroacetic acid) to give0.12 g (92%) of the trifluoroacetic acid salt of the title compound as apale yellow foam. MS (ES⁺): (M+H)=717.5

EXAMPLE 35N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following General Procedure I Method C,N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(0.267 g, 0.565 mmol) and L-proline methylester hydrochloride salt(0.112 g, 0.678 mmol, Fluka) are coupled to give the trifluoroaceticacid salt of the title compound as a yellow oil, 0.17 g (51% yield), MSm/z 585.3 (M+H).

EXAMPLE 36N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following General Procedure IIN,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(0.146 g, 0.25 mmol) from Example 35 is hydrolyzed and the productisolated by HPLC (0.01% trifluoroacetic acid in water/acetonitrile) togive the trifluoroacetic acid salt of the title compound as whitecrystals, 0.014 g. MS: m/z 571.4 (M+H)

EXAMPLE 37N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(2-phenethoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamid

To a solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(0.089 g, 0.156 mmol, from Example 36) hydroxybenzotriazole (0.025 g,0.187 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiiminehydrochloride (0.042 mg, 0.22 mmol) in anhydrous dimethylformamide (0.5ml) is added diisopropylethylamine (0.054 ml, 0.312 mmol) under an inertatmosphere. To this solution is added a solution of phenethylamine(0.023 g, 0.187 mmol) in anhydrous dimethylformamide (0.5 ml). Theresulting reaction mixture is stirred at room temperature overnight. Thesolvent is removed in vacuo. The residue is chromatographed by HPLC(0.01% trifluoroacetic acid in water/acetonitrile) to give thetrifluoroacetic acid salt of the title compound as white crystals, 0.052g (50% yield). MS: m/z 674.6 (M+H)

EXAMPLE 38N,β,β-trimethyl-L-phenylalanyl-N¹-((1S,2E)-4-{4-[3-(dimethylamino)propyl]piperazin-1-yl}-1-isopropyl-3-methyl-4-oxobut-2-enyl)-N¹,3-dimethyl-L-valinamide

Following General Procedure I Method C,N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(0.190 g, 0.402 mmol) and 1-(3-dimethylaminopropyl)-piperazine (0.0825g, 0.482 mmol) are coupled to give the trifluoroacetic acid salt of thetitle compound as a white solid, 0.091 g (36% yield), MS m/z 627.6 (M+H)

EXAMPLE 39N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-4-(4-benzylpiperazin-1-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

Following General Procedure I Method C,N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(0.239 g, 0.505 mmol) and 1-benzylpiperazine (0.107 g, 0.606 mmol,Aldrich) are coupled to give the trifluoroacetic acid salt of the titlecompound as a white crystals, 0.378 g, MS m/z 632.5 (M+H)

EXAMPLE 40N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S,4R)-2-(methoxycarbonyl)-4-hydroxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following General Procedure I Method C,N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(0.270 g, 0.571 mmol) and L-trans-4-hydoxyproline methyl esterhydrochloride salt (0.099 g, 0.685 mmol, Advance ChemTech) are coupledto give the trifluoroacetic acid salt of the title compound as a whitesolid 0.2162 g (62% yield), MS m/z 601.4 (M+H)

EXAMPLE 41N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2R)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following General Procedure I Method C,N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(0.224 g, 0.47 mmol) and D-proline methyl ester hydrochloride salt(0.094 g, 0.57 mmol) are coupled to give the trifluoroacetic acid saltof the title compound as a white solid 0.196 g (58% yield), MS m/z 585.5(M+1)

EXAMPLE 42N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S,4R)-2-carboxy-4-hydroxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following General Procedure IIN,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S,4R)-2-(methoxycarbonyl)-4-hydroxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(0.133 g, 0.221 mmol) from Example 40 is hydrolyzed and the productisolated by HPLC (0.01% trifluoroacetic acid in water/acetonitrile) togive the trifluoroacetic acid salt of the title compound as a lightyellow solid, 0.078 g (60% yield), MS: m/z 585.3 (M−H)

EXAMPLE 43N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2R)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following General Procedure II,N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2R)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(0.159g, 0.273 mmol) from Example 41 is hydrolyzed and the productisolated by HPLC (0.01% trifluoroacetic acid in water/acetonitrile) togive the trifluoroacetic acid salt of the title compound as a whitesolid, 0.138 g (88% yield), MS: m/z 569.4 (M−1)

EXAMPLE 443-cyclohexyl-N-methyl-L-valyl-N¹-{(1S,2E)-1-isopropyl-4-[(2S)-2-((1R,2R)-1methoxy-2-methyl-3-oxo-3-{[(1S)-2-phenyl-1-(1,3-thiazol-2-yl)ethyl]amino}propyl)pyrrolidin-1-yl]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following General Procedure I Method C,3-cyclohexyl-N-methyl-L-valyl-N-1-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N-1,3-dimethyl-L-valinamide(0.083 g, 0.173 mmol) and (2R,3R,4S)-dolaproine-(S)-dolaphenine (0.077g, 0.208 mmol, from Example 29) are coupled to give the trifluoroaceticacid salt of the title compound as a white solid 0.064 g (44% yield), MSm/z 835.7 (M+H)

EXAMPLE 453-cyclohexyl-N-methyl-L-valyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following General Procedure I Method C,3-cyclohexyl-N-methyl-L-valyl-N-1-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N-1,3-dimethyl-L-valinamide(0103 g, 0.215 mmol) and L-proline methyl ester hydrochloride salt(0.043 g, 0.257 mmol, Aldrich) are coupled to give the trifluoroaceticacid salt of the title compound as a light yellow solid 0.068 g (53%yield), MS m/z 591.4 (M+1)

EXAMPLE 463-cyclohexyl-N-methyl-L-valyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following the General Procedure II3-cyclohexyl-N-methyl-L-valyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(0.091 g, 0.154 mmol, from Example 45) is hydrolyzed and the productisolated by HPLC (0.01% trifluoroacetic acid in water/acetonitrile) togive the trifluoroacetic acid salt of the title compound as white solid,0.070 g (79% yield), MS: m/z 575.5 (M−H)

EXAMPLE 47N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[(2S)-2-(methoxycarbonyl)-2,5-dihydro-1H-pyrrol-1-yl]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

To the solution of BOC-3,4-dehydro-L-proline (1.03 g, 4.84 mmol) indiethylether (3 ml) at room temperature is added(trimethylsilyl)diazomethane (6.8 ml, 13.7 mmol). The reaction mixtureis stirred at room temperature for 35 minute. Solvent is removed invacuo. The residue is purified by preparative TLC (2000 micron silicaplate, eluted with 3.5:1 hexane/ethyl acetate) to giveBOC-3,4-dehydro-L-proline-methyl ester as a colorless oil 0.426 g. Thenthis oil is treated with 4 N hydrogen chloride in dioxane (11.3 ml,27.62 mmol) to give 3,4-dehydro-L-proline methyl ester hydrochloridesalt. Following General Procedure I Method C,4-[[(2S)-3,3-Dimethyl-2-[[(2S)-3-methyl-2-(methylamino)-1-oxo-3-phenylbutyl]amino)]-1-oxobutyl]methylamino]-2,5-dimethyl-(E,4S)-2-hexenoic acid (0.672 g, 1.42 mmol) and 3,4-dehydro-L-prolinemethyl ester hydrochloride salt (0.281 g, 1.71 mmol,) are coupled togive the trifluoroacetic acid salt of the title compound as a whitesolid 0.608 g (73% yield), MS m/z 583.4 (M+1)

EXAMPLE 48N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxy-2,5-dihydro-1H-pyrrol-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following General Procedure I Method B,4-[[(2S)-3,3-Dimethyl-2-[[(2S)-3-methyl-2-(methylamino)-1-oxo-3-phenylbutyl]amino)]-1-oxobutyl]methylamino]-2,5-dimethyl-(E,4S)-2-hexenoic acid (0.262 g, 0.55 mmol) and 3,4-dehydro-L-prolinemethyl ester hydrochloride salt (0.099 g, 0.66 mmol,) are coupled togive the trifluoroacetic acid salt of the title compound as a whitesolid 0.196 g (58% yield), MS m/z 569.3 (M+1)

EXAMPLE 49N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[(2S)-2-(methoxycarbonyl)piperidin-1-yl]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following General Procedure I Method C,4-[[(2S)-3,3-Dimethyl-2-[[(2S)-3-methyl-2-(methylamino)-1-oxo-3-phenylbutyl]amino)]-1-oxobutyl]methylamino]-2,5-dimethyl-(E,4S)-2-hexenoic acid (0.476 g, 1.00 mmol) and L-pipecolic acid methylester hydrochloride salt (0.099 g, 0.66 mmol,) are coupled to give thetrifluoroacetic acid salt of the title compound as a white solid 0.514 g(85% yield), MS m/z 599.3 (M+1)

EXAMPLE 50N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypiperidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following the General Procedure II,N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[(2S)-2-(methoxycarbonyl)piperidin-1-yl]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(0.241 g, 0.402 mmol) from Example 49 is hydrolyzed and the productisolated by HPLC (0.01% trifluoroacetic acid in water/acetonitrile) togive the trifluoroacetic acid salt of the title compound as white solid,0.232 g (99% yield), MS: m/z 585.3 (M+H)

EXAMPLE 513-cyclohexyl-N-methyl-L-valyl-N¹-[(1S,2E)-1-isopropyl-4-((2S)-2-{(1R,2R)-1-meyhoxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidin-1-yl)-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamide

Following the diethyl cyanophosphonate coupling procedure described inExample 29,3-cyclohexyl-N-methyl-L-valyl-N-1-[(1S,2E)-3-carboxy-1-isopropyl-2-butenyl]-N-1,3-dimethyl-L-valinamide(0.135 g, 0.282 mmol) and(2S)-2-{(1R,2R)-1-methoxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidine(0.117 g, 0.34 mmol, from Example 32) are coupled to give the titlecompound as a white solid 0.041 g (18% yield), MS m/z 808.4 (M+1)

EXAMPLE 52N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[methoxy(methyl)-amino]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

A solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(473 mg, 1 mmol, WO 99/32509) in tetrahydrofuran (2 mL) is added asolution of N,O-dimethylhydroxylamine hydrochloride (166 mg, 1.7 mmol)in water (2 mL), adjusting the reaction mixture to pH 4.5 by addingdropwise 1.0 N sodium hydroxide solution. To this is added dropwise asolution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimine hydrochloride(479 mg, 2.5 mmol) in water (7 mL) over a period of 7 min, maintainingpH 4.5 by using 1.0 N sodium hydroxide solution. The reaction mixture isstirred at room temperature for 2.5 h, adjusting the pH to 6-7. To thisis added brine (10 mL) and extracted with dichloromethane. The combinedorganic layers are washed with brine, dried (Na₂SO₄), filtered andconcentrated to give 491 mg (95%) of the title compound as a whitesolid. HRMS (ESI) calcd for C₂₉H₄₈N₄O₄ (M+H⁺) 517.3748, found 517.3750.

EXAMPLE 53N,β,β-trimethyl-L-phenylalanyl-N-1-{(1S,2E)-4-[(1,1′-biphenyl-4-ylmethyl)amino]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide

Following General Procedure I Method AN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(256 mg, 0.542 mmol, WO 99/32509) and 4-phenylbenzylamine (119 mg, 0.65mmol) are coupled to give the title compound as a yellow oil, 83 mg, MS:m/z 639.4 (M+1).

EXAMPLE 54N,β,β-trimethyl-L-phenylalanyl-N¹-((1R,2E)-4-{[4-(benzyloxy)benzyl]oxy}-1-isopropyl-3-methyl-4-oxobut-2-enyl)-N¹,3-dimethyl-L-valinamide

A mixture ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(287 mg, 0.607 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiiminehydrochloride (0.728 mmol), 4-(benzyloxy)benzyl alcohol (137 mg, 0.638mmol) and dimethylaminopyridine (0.12 mmol) in anhydrous dichloromethane(12 mL) are stirred under a nitrogen atmosphere at room temperature for20 hours. The solvent is removed, the residue taken up in methanol, andthe product purified by reverse phase HPLC (0.01% trifluoroacetic acidin water/acetonitrile) to give the title compound as a white gum, 208mg, MS: m/z 670.4 (M+1).

REFERENCE EXAMPLE 6 Adamantan-1-yl-methylamino-acetic acid

Adamantan-1-yl-iodo-acetic acid (7.86 g, 24.55 mmol, P. A. Krasutskii.et. Al.;J. Org. Chem. USSR (Engl. Trans.); 1985; 1327-1332) is refluxedwith 100 mL of 2M methylamine in tetrahydrofuran for 18 hours. Thesolvent is evaporated in vacuo, and the residue treated with water. ThepH is adjusted to 6 and the resulting solid collected and dried to give2.16 g of the title compound as a white solid. MS: m/z 224.2 (M+1).

REFERENCE EXAMPLE 7 Ethyl(2E,4S)-4-[{N-[(2S)-2-(1-adamantyl)-2-(methylamino)ethanoyl]-3-methyl-L-valy}(methyl)amino]-2,5-dimethyl-2-hexenoateand REFERENCE EXAMPLE 8 Ethyl(2E,4S)-4-[{N-[(2R)-2-(1-adamantyl)-2-(methylamino)ethanoyl]-3-methyl-L-valy}(methyl)amino]-2,5-dimethyl-2-hexenoate

To adamantan-1-yl-methylamino-acetic acid (0.447 g, 2.0 mmol, fromReference Example 6) in dichloromethane (17 mL) is added PyBOP (1.04 g,2.0 mmol). After 15 min ethyl(2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoate(0.625 g, 2.0 mmol, WO 99/32509) in dichlormethane (5 mL) is added,followed by diisopropylethylamine (0.75 mL, 4.3 mmol). This is stirredat room temperature overnight. The reaction mixture is evaporated invacuo and the residue treated with ethyl acetate and water. The ethylacetate layer is dried over sodium sulfate and purified by hplc. The twodiastereomers are partially separated. The first isomer is assigned theSSS configuration and contained 33.4% of the RSS isomer. MS: m/z 518.3(M+1). The second isomer is assigned the RSS configuration and contained8.9% of the SSS isomer. MS: m/z 518.3 (M+1).

EXAMPLE 55(2E,4S)-4-[{N-[(2S)-2-(1-adamantyl)-2-(methylamino)ethanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoicacid

According to General Procedure II ethyl(2E,4S)-4-[{N-[(2S)-2-(1-adamantyl)-2-(methylamino)ethanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoate(303 mg, 0.59 mmol, from Reference Example 7) in water (1.5 mL) andmethanol (6.0 mL) is treated with aqueous 1 M lithium hydroxide solution(2.9 mL, 2.9 mmol) to provide the trifluoroacetic acid salt of the titlecompound (49 mg) as a white solid after preparative HPLC. MS (ES): m/z490.24 (M+H).

EXAMPLE 56(2E,4S)-4-[{N-[(2R)-2-(1-adamantyl)-2-(methylamino)ethanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoicacid

According to General Procedure II ethyl(2E,4S)-4-[{N-[(2R)-2-(1-adamantyl)-2-(methylamino)ethanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoate(359 mg, 0.69 mmol, from Reference Example 8) in water (1.7 mL) andmethanol (7.0 mL) is treated with aqueous 1 M lithium hydroxide solution(3.5 mL, 3.5 mmol) to provide the trifluoroacetic acid salt of the titlecompound (62 mg) as a white solid after preparative HPLC. MS (ES): m/z490.24 (M+H).

EXAMPLE 57(2E,4S)-2,5-dimethyl-4-(methyl{3-methyl-N-[(2S)-2-(methylamino)-2-phenylethanoyl]-L-valyl}amino)-2-hexenoicacid

To N-methyl phenylglycine (330 mg, 2.0 mmol) in dichloromethane (15 mL)is added PyBOP (1.04 g, 2.0 mmol). After 15 mins, ethyl(2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoate(0.624 g, 2.0 mmol, WO 99/32509) in dichlormethane (5 mL) is addedfollowed by diisopropylethylamine (0.8 mL, 4.4 mmol). After 18 h thereaction mixture is concentrated in vacuo and the residue treated withethyl acetate and water. The ethyl acetate layer is dried over sodiumsulfate, filtered and concentrated to give 772 mg of a white solid, MS:m/z 460.46 (M+1). According to General Procedure II this solid (689 mg,1.5 mmol) in water (3.6 mL) and methanol (15 mL) is treated with aqueous1 M lithium hydroxide solution (7.5 mL, 7.5 mmol), to provide thetrifluoroacetic acid salt of the title compound (541 mg) as a whitesolid after preparative HPLC. MS (ES): m/z 432.42 (M+H).

EXAMPLE 58(2E,4S)-2,5-dimethyl-4-(methyl{3-methyl-N-[(2,2,4-trimethylthiomorpholin-3-yl)carbonyl]-L-valyl}amino)hex-2-enoicacid

A mixture of 2,2-Dimethyl-thiomorpholine-3-carboxylic acid (3.51 g, 20mmol, BE 893025), formic acid (5.12 g of 90%) and formalin (4.5 mL of37%) is heated at reflux for 8 hours. The reaction mixture is thencooled and treated with hydrochloric acid (10 mL of 4N) and evaporatedto dryness. A portion of this residue is purified by HPLC to give2,2,4-trimethyl-thiomorpholine-3-carboxylic acid, which is used as is.2,2,4-Trimethyl-thiomorpholine-3-carboxylic acid (388 mg, 2.05 mmol) isstirred with dichloromethane (4 mL) and dimethylformamide (4 mL). Tothis is added (benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate (PyBOP, 1.07 g, 2.05 mmol). After 15 min, ethyl(2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoate(0.640 g, 2.05 mmol, WO 99/32509) is added in dichlormethane (5 mL)followed by diisopropylethylamine (0.8 mL, 4.4 mmol). This is stirred atroom temperature overnight. The reaction mixture is evaporated in vacuoand the residue treated with ethyl acetate and water. The ethyl acetatelayer is dried over sodium sulfate, filtered and concentrated. Theresulting material (314 mg, 0.65 mmol) is treated with aqueous 1 Mlithium hydroxide solution (3.3 mL, 3.3 mmol), water (1.6 mL) andmethanol (6.5 mL), to provide the trifluoroacetic acid salt of the titlecompound as a white solid after preparative HPLC, MS (ES): m/z 456.29(M+H).

REFERENCE EXAMPLE 9 Ethyl (2Z)-3,4-dihydro-1(2H)-naphthalenylidene(formylamino)ethanoate

To a stirred slurry of potassium t-butoxide (4.7 g, 42.0 mmol) in 30 mLtetrahydrofuran at 0° C. is added ethyl isocyanoacetate (4.53 g, 40.0mmol). This is stirred for 30 mins whereupon a solution of β-tetralone(5.88 g, 40.0 mmol) in tetrahydrofuran (20 mL) is added dropwise. Thereaction mixture is allowed to warm to room temperature and stirred for3 hours. The reaction mixture is then recooled in an ice bath and tothis is added acetic acid (2.5 g, 41.6 mmol), allowed to warm to roomtemperature and stirred for an additional 30 mins. The solvent isremoved in-vacuo and the residue partitioned between water and ethylacetate. The organic layer is washed with water and brine, dried oversodium sulfate, filtered, evaporated and crystallized from cyclohexaneto give 4.67 g of a white solid (contains some E-isomer), MS: m/z 260.2(M+1).

REFERENCE EXAMPLE 10 Methyl(1-methyl-1,2,3,4-tetrahydro-1-naphthalenyl)(oxo)acetate

Ethyl (2Z)-3,4-dihydro-1(2H)-naphthalenylidene(formylamino)ethanoate(1.31 g, 5.05 mmol, from Reference Example 9) is refluxed with 6N HCL(10 mL) for 1 hour. The mixture is evaporated, dried and the residuetaken up in tetrahydrofuran (8 mL) and water (1.7 mL). To this is added5 N sodium hydroxide solution (3.6 mL, 18.0 mmol) and iodomethane (0.9mL, 14.5 mmol). While stirring under a nitrogen atmosphere, the reactionmixture is heated in a 70° C. oil bath for 5½ hours. Additional volumesof 5 N sodium hydroxide solution (2.0 mL) and iodomethane (0.3 mL, 4.8mmol) are added. The reaction mixture is allowed to stir for 2 days atroom temperature, then evaporated in vacuo to remove most of thetetrahydrofuran. This is extracted with ethyl acetate and the organiclayer then dried over anhydrous sodium sulfate, filtered, andconcentrated in vacuo to give a clear oil, 0.49 g, MS: m/z 233.2 (M+1).

EXAMPLE 59(2E,4S)-2,5-dimethyl-4-(methyl{3-methyl-N-[(2S)-2-(methylamino)-2-(1-methyl-1,2,3,4-tetrahydro-1-naphthalenyl)ethanoyl]-L-valyl}amino)-2-hexenoicacid and EXAMPLE 60(2E,4S)-2,5-dimethyl-4-(methyl{3-methyl-N-[(2R)-2-(methylamino)-2-(1-methyl-1,2,3,4-tetrahydro-1-naphthalenyl)ethanoyl]-L-valyl}amino)-2-hexenoicacid

Methyl (1-methyl-1,2,3,4-tetrahydro-1-naphthalenyl)(oxo)acetate (0.400g, 1.72 mmol, from Reference Example 10) is treated with 1 M lithiumhydroxide solution (6.9 mL, 6.9 mmol) and methanol (15 mL) and stirredovernight. The reaction mixture is then acidified and extracted withethyl acetate, dried over sodium sulfate, filtered and evaporated. Theproduct is dissolved in tetrahydrofuran (4 mL) and treated withmethylamine (4.0 mL of 2M in tetrahydrofuran, 8.0 mmol) and stirred for1 hour then a solution of borane-pyridine complex (0.26 mL of 8Msolution, 2.08 mmol) is added. The mixture is heated at 50° C. for 2.5hours and cooled to room temperature. Methanol (5 mL) is added and thereaction mixture then evaporated to dryness and triturated withtetrahydrofuran to give a white solid. This solid (116 mg, 0.50 mmol) isstirred with dichloromethane (2 mL) and dimethylformamide (2 mL). Tothis is added PyBOP (0.260 g, 0.50 mmol) and stirred for 15 mins, theBCD ester (0.156 g, 0.50 mmol) is added in dichlormethane (1 mL)followed by diisopropylethylamine (0.2 mL, 1.1 mmol). This is stirred atroom temperature overnight. The reaction mixture is evaporated in vacuoand the residue treated with ethyl acetate and water. The ethyl acetatelayer is dried over sodium sulfate, filtered and concentrated giving theester. This ester (169 mg, 0.32 mmol) is treated with aqueous 1M lithiumhydroxide solution (1.6 mL, 1.6 mmol), water (0.8 mL) and methanol (3.2mL), to provide after preparative HPLC two components. The first isomereluted is arbitrarily assigned as the S-isomer (Example 59) (11 mg of awhite solid), MS (ES): m/z 500.32 (M+H). The second component (Example60) is 43 mg of a white solid, MS (ES): m/z 500.33 (M+H), which has 27%of Example 59 present.

REFERENCE EXAMPLE 11Ethyl-3-hydroxy-L-valyl-N¹-[(1S,2E)-4-ethoxy-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹-L-valinamide

To a cooled (0° C., ice bath) solution of(S)-(+)-2-amino-3-hydroxy-3-methylbutanoic acid (42.7 mg, 0.32 mmol) andbenzotriazole-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate(192.3 mg, 0.37 mmol) in anhydrous dimethylformamide (3-5 ml) is addeddiisopropylethylamine (0.167 ml, 0.96 mmol). To this solution is added asolution of ethyl(E,4S)-4-[[(2S)-2-amino-3,3-dimethylbutanoyl](methyl)amino]-2,5-dimethyl-2-hexenoate(0.10 g, 0.32 mmol, WO 99/32509) in anhydrous dimethylformamide (3 ml).After stirring at 0° C. for 5-10 minutes, the cooling bath is removed,and the resulting reaction mixture is stirred at room temperature for15-20 hours. The mixture is diluted with water, and the aqueous layer isextracted with ethyl acetate (3 times). The combined extracts are washedwith saturated aqueous sodium chloride, dried over sodium sulfate,filtered and concentrated in vacuo. The residue is chromatographed(silica gel, flash column), to give the title compound as a white solid.(70 mg, 76%). MS (ES) m/z 425.29 [M+H]

EXAMPLE 613-hydroxy-N-methyl-L-valyl-N¹-[(1S,2E)-4-ethoxy-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamideand3-hydroxy-N,N-dimethyl-L-valyl-N¹-[(1S,2E)-4-ethoxy-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamide

To a vigorously stirred solution ofethyl-3-hydroxy-L-valyl-N¹-[(1S,2E)-4-ethoxy-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹-L-valinamide(50 mg, 0.12 mmol, from Reference Example 11) in anhydrousdimethylformamide (1 ml) at 0° C. (ice bath) is added iodomethane (0.017ml, 0.12 mmol). After 30 minutes diisopropylethylamine (0.042 ml, 0.24mmol) is added dropwise. The reaction mixture is allowed to warm to roomtemperature. After 2 hours water is added and the mixture is acidifiedby dropwise addition of 1.0M aqueous citric acid. The acidic mixture isextracted 3× with ethyl acetate. The combined organic layers are driedover sodium sulfate and concentrated in vacuo to give a mixture of ethyl3-hydroxy-N-methyl-L-valyl-N¹-[(1S,2E)-4-ethoxy-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamidecompound andethyl-3-hydroxy-N,N-dimethyl-L-valyl-N¹-[(1S,2E)-4-ethoxy-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamide.This material (50 mg) is dissolved in methanol (1 ml) andtetrahydrofuran (1 ml). To the resulting solution is added 2.0 M aqueouslithium hydroxide solution (0.1 mL). After 15 hours the volatiles areremoved in vacuo, and the residual aqueous mixture is cooled with an icewater bath, and acidified to pH 5.5-6.0 with aqueous 1N aqueous citricacid solution. The resulting precipitate is collect by filtration, andthe solid is washed with a cold water, and dried over high vacuum. Thecrude material is purified by reverse phase HPLC to give thetrifluoroacetic acid salt of the title compounds as a white solid (40mg, 80%). A 1:1 mixture of components is present by analytical HPLC/MS:m/z3-hydroxy-N-methyl-L-valyl-N¹-[(1S,2E)-4-ethoxy-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamide:[M+H] 414.33-hydroxy-N,N-dimethyl-L-valyl-N¹-[(1S,2E)-4-ethoxy-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamide:[M+H] 428.4

EXAMPLE 621-methyl-L-prolyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide

To a solution of N-methyl-L-proline monohydrate (0.26 g, 1.79 mmol) andbenzotriazole-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate(1.4 g, 2.69 mmol) in anhydrous dichloromethane (3.0 ml) at 25° C. isadded diisopropylethylamine (0.63 ml, 3.62 mmol) under an inertatmosphere. To this solution is added a solution of ethyl(2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoate (0.39g, 1.27 mmol, WO 99/32509) in anhydrous dichloromethane (2.0 ml). Theresulting reaction mixture is stirred at room temperature overnight. Thereaction mixture is evaporated in vacuo. The residue (300 mg, 0.78 mmol)is dissolved in methanol (6.85 ml), water (1.8 ml) and tetrahydrofuran(3.25 ml). To this reaction mixture is added aqueous lithium hydroxidesolution (3.54 ml, 3.54 mol) The resulting mixture is stirred at roomtemperature overnight. The reaction mixture is diluted withdichloromethane/water. The organic layer is washed with citric acid,brine, dried over sodium sulfate and filtered. The solvent is removed invacuo, and the residue is chromatographed by HPLC (0.01% trifluoroaceticacid in water/acetonitrile) to give the product as a yellow oil, 0.08 g(28% yield). MS: m/z 396.28 (M+1)

REFERENCE EXAMPLE 12N-{1-[(1-Formyl-2-methyl-propyl)-methyl-carbamoyl]-2,2-dimethyl-propyl}-3-methyl-2-methylamino-3-phenyl-butyramide

A solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(1.0 g) in methanol (20 mL) is added to 100 mL of methanol at −78° C.which had been treated with ozone until the solution is blue. Theresulting colorless reaction mixture is treated with ozone at −78° C.until the blue color remained. After 5 min dimethylsulfide (2 mL) isadded. The reaction mixture is allowed to warm to room temperature andthen concentrated in vacuo to give the title compound as an oil. MS(ES): m/z 418.4 (M+H).

EXAMPLE 63N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(ethoxysulfonyl)-1-isopropylprop-2-enyl]-N¹,3-dimethyl-L-valinamide

To a solution of ethyl diethylphosphorylmethanesulfonate (260 mg, 1.0mmol) in anhydrous THF (3 mL) at −78° C. is added n-butyllithium (inhexanes, 0.44 mL, 1.1 mmol). After stirring for 20 min at −78° C., asolution ofN-{1-[(1-Formyl-2-methyl-propyl)-methyl-carbamoyl]-2,2-dimethyl-propyl}-3-methyl-2-methylamino-3-phenyl-butyramide(1.0 mmol) in anhydrous THF (7 mL), which is freshly prepared asdescribed in Reference Example 12, is added. The reaction mixture isstirred at −78° C. for additional 30 min, and then gradually warmed toroom temperature. After 40 h, the reaction is quenched with water andextracted with CH₂Cl₂. The combined organic layers are washed withbrine, dried (Na₂SO₄), filtered and concentrated. The residue ischromatographed on a silica gel column, eluting with 4:1 CH₂Cl₂/EtOAc togive 160 mg (31%) of the title compound as a yellow oil. HRMS (ESI)calcd for C₂₇H₄₅N₃O₅S [M+H] 524.3153, found 524.3157.

EXAMPLE 64N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-sulfoprop-2-enyl]-N¹,3-dimethyl-L-valinamide

A solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(ethoxysulfonyl)-1-isopropylprop-2-enyl]-N¹,3-dimethyl-L-valinamide(30 mg, 0.057 mmol) from Example 63 in acetone (1 mL) is treated withtetrabutylammonium iodide (21 mg, 0.057 mmol). The reaction mixture isrefluxed for 9 h, and then stirred at room temperature overnight. Thesolvent is removed. The residue is purified by reversal phase HPLC(mobile phase A: 0.1% TFA/5% acetonitrile/H₂O, mobile phase B: 100%acetonitrile) to give 10 mg (28%) of the title compound as a whitesolid. HRMS (ESI) calcd for C₂₅H₄₁N₃O₅S [M+H] 496.2840, found 498.2840.

EXAMPLE 65N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(ethoxysulfonyl)-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide

To a solution of ethyl 1-(diethoxyphosphoryl)ethanesulfonate (210 mg,0.77 mmol) in anhydrous THF (4 mL) cooled to −78° C. is addedbutyllithium (0.4 mL, 1.0 mmol). After stirring for 20 min at −78° C., asolution ofN-{1-[(1-Formyl-2-methyl-propyl)-methyl-carbamoyl]-2,2-dimethyl-propyl}-3-methyl-2-methylamino-3-phenyl-butyramide(0.77 mmol, from Reference Example 12) in anhydrous THF (4 mL), isadded. The reaction mixture is stirred at −78° C. for an additional 3 h,and then gradually warmed to room temperature overnight. The reaction isquenched with water, extracted with CH₂Cl₂. The combined organic layersare washed with brine, dried (Na₂SO₄), filtered and concentrated. Theresidue is separated by reversal phase HPLC (mobile phase A: 0.1% TFA/5%acetonitrile/H₂O, mobile phase B: 100% acetonitrile) to give 97 mg (19%)of white solids which contains the trifluoroacetic acid salt of thetitle compound along with a small amount of its diastereoisomers. HRMS(ESI) calcd for C₂₈H₄₇N₃O₅S [M+H] 538.3309, found 538.3302.

EXAMPLE 66N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-sulfobut-2-enyl]N¹,3-dimethyl-L-valinamide and EXAMPLE 67N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2Z)-1-isopropyl-3-sulfobut-2-enyl]-N¹,3-dimethyl-L-valinamide

A solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(ethoxysulfonyl)-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(30 mg, 0.046 mmol, from Example 65) in acetone (2 mL) is treated withtetrabutylammonium iodide (20 mg, 0.055 mmol). The reaction mixture isrefluxed for 7 h, and then stirred at room temperature overnight. Thesolvent is removed. The residue is purified by reversal phase HPLC(mobile phase A: 0.1% TFA/5% acetonitrile/H₂O, mobile phase B: 100%acetonitrile) to give 12 mg (28%) ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-sulfobut-2-enyl]N¹,3-dimethyl-L-valinamide as a white solid and 2 mg ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2Z)-1-isopropyl-3-sulfobut-2-enyl]-N¹,3-dimethyl-L-valinamide

-   HRMS (ESI) for    N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-sulfobut-2-enyl]-N¹,3-dimethyl-L-valinamide,    calcd for C₂₆H₄₃N₃O₅S [M+H] 510.2996, found 510.2986.-   HRMS (ESI) for    N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2Z)-1-isopropyl-3-sulfobut-2-enyl]-N¹,3-dimethyl-L-valinamide,    calcd for C₂₆H₄₃N₃O₅S [M+H] 510.2996, found 510.2991.

EXAMPLE 68N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(dimethoxyphosphoryl)-1-isopropylprop-2-enyl]-N¹,3-dimethyl-L-valinamide

To a solution of tetramethyl methylenediphosphonate (97 mg, 0.42 mmol)in anhydrous THF (2 mL) cooled to −78° C. is added butyllithium (0.17mL, 0.42 mmol). After stirring for 20 min at −78° C., a solution ofN-{1-[(1-Formyl-2-methyl-propyl)-methyl-carbamoyl]-2,2-dimethyl-propyl}-3-methyl-2-methylamino-3-phenyl-butyramide(100 mg, 0.21 mmol, from Reference Example 12) in anhydrous THF (1 mL),is added. The reaction mixture is stirred at −78° C. for additional 30min, and then gradually warmed to room temperature for 40 h. Thereaction is quenched with water, extracted with CH₂Cl₂. The combinedorganic layers are washed with brine, dried (Na₂SO₄), filtered andconcentrated. The residue is purified by reversal phase HPLC (mobilephase A: 0.1% TFA/5% acetonitrile/H₂O, mobile phase B: 100%acetonitrile) to give 30 mg (22%) of the trifluoroacetic acid salt ofthe title compound as a white solid. HRMS (ESI) calcd for C₂₇H₄₆N₃O₅P[M+H] 524.3248, found 524.3244.

EXAMPLE 69N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-3-[hydroxy(methoxy)phosphoryl]-1-isopropylprop-2-enyl}-N¹,3-dimethyl-L-valinamide

A solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(dimethoxyphosphoryl)-1-isopropylprop-2-enyl]-N¹,3-dimethyl-L-valinamide(7 mg, 0.013 mmol, from Example 68) in CH₂Cl₂ (0.3 mL) is treated withTMSBr (3.5 mg, 0.023 mmol). The reaction mixture is stirred at roomtemperature for 2 h, then concentrated. The residue is purified byreverse phase HPLC (mobile phase A: 0.1% TFA/5% acetonitrile/H₂O, mobilephase B: 100% acetonitrile) to give 2 mg (24%) of the trifluoroaceticacid salt of the title compound as a white solid. HRMS (ESI) calcd forC₂₆H₄₄N₃O₅P [M−H] 508.2946, found 508.2939.

EXAMPLE 70N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-phosphonoprop-2-enyl]N¹,3-dimethyl-L-valinamide

A solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(dimethoxyphosphoryl)-1-isopropylprop-2-enyl]-N¹,3-dimethyl-L-valinamide(7 mg, 0.013 mmol, from Example 68) in CH₃CN (0.3 mL) is treated withTMSBr (6.1 mg, 0.04 mmol). The reaction mixture is stirred at roomtemperature for 4 h. Additional TMSBr (6.1 mg) is added. The reactionmixture is stirred for additional 4 h, then concentrated. The residue ispurified by reverse phase HPLC (mobile phase A: 0.1% TFA/5%acetonitrile/H₂O, mobile phase B: 100% acetonitrile) to give 2 mg (25%)of the trifluoroacetic acid salt of the title compound as a white solid.HRMS (ESI) calcd for C₂₅H₄₂N₃O₅P [M−H] 494.2789, found 494.2790.

EXAMPLE 71N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(diethoxyphosphoryl)-1-isopropyl-but-2-enyl]-N¹,3-dimethyl-L-valinamide

To a solution of tetraethyl ethyl-1,1-bisphosphonate (65 mg, 0.215 mmol)in anhydrous THF (1 mL) cooled to −78° C. is added butyllithium (0.11mL, 0.268 mmol). After stirring for 20 min at −78° C., a solution ofN-{1-[(1-Formyl-2-methyl-propyl)-methyl-carbamoyl]-2,2-dimethyl-propyl}-3-methyl-2-methylamino-3-phenyl-butyramide(100 mg, 0.215 mmol, from Reference Example 12), in anhydrous THF (1mL), is added. The reaction mixture is stirred at −78° C. for anadditional 30 min, and then gradually warmed to room temperature for 60h. The reaction is quenched with water, extracted with CH₂Cl₂. Thecombined organic layers are washed with brine, dried (Na₂SO₄), filteredand concentrated. The residue is purified by reversal phase HPLC (mobilephase A: 0.1% TFA/5% acetonitrile/H₂O, mobile phase B: 100%acetonitrile) to give 16 mg (11%) of a mixture of the major desiredproduct along with small amount of its diastereoisomers as a whitesolid. HRMS (ESI) calcd for C₃₀H₅₂N₃O₅P [M+H] 566.3717, found 566.3720.

EXAMPLE 72N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-phosphonobut-2-enyl]-N¹,3-dimethyl-L-valinamide

A solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-(diethoxyphosphoryl)-1-isopropyl-but-2-enyl]-N¹,3-dimethyl-L-valinamide(11 mg, 0.02 mmol, from Example 71) in CH₂Cl₂ (0.5 mL) is treated withTMSBr (23 mg, 0.15 mmol). The reaction mixture is stirred at roomtemperature for 20 h, then concentrated. The residue is purified byreversal phase HPLC (mobile phase A: 0.02% TFA/H₂O, mobile phase B:0.02% TFA/CH₃CN) to give 2.4 mg (19%) of desired product as a whitesolid. HRMS (ESI) calcd for C₂₆H₄₄N₃O₅P [M+H] 510.3091, found 510.3086.

EXAMPLE 73N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-4-(1,3-thiazol-2-yl)but-2-enyl]-N¹,3-dimethyl-L-valinamide

To a solution of thiazole (0.035 mL, 0.5 mmol) and TMEDA (0.068 mL, 0.45mmol) in anhydrous THF (1.0 mL) cooled to −60° C. is added n-BuLi (0.17mL, 0.43 mmol). After stirring for 20 min at −60° C., a solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[methoxy(methyl)-amino]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(52 mg, 0.1 mmol, from Example 52, in anhydrous THF (0.5 mL) is added.The purple reaction mixture is stirred for 2 h from −60° C. to roomtemperature, quenched with saturated aqueous ammonium chloride (1.5 mL).The reaction mixture is extracted with ethyl acetate three times. Thecombined organic layers are dried (Na₂SO₄), filtered and concentrated.The residue is purified by reversal phase HPLC (mobile phase A: 0.1%TFA/5% acetonitrile/H₂O, mobile phase B: 100% acetonitrile) to give 56mg (86%) of desired product as a yellow solid.

HRMS (ESI) calcd for C₃₀H₄₄N₄O₃S [M+H] 541.3207, found 541.3203.

EXAMPLE 74N,β,β-trimethyl-L-phenylalanyl-N-1-[(1S,2E)-4-hydroxy-1-isopropyl-3-methyl-4-phenylbut-2-enyl}-N-1,3-dimethyl-L-valinamide

To a slurry of lithium aluminum hydride (12 mg, 0.32 mmole) in ether (5mL) cooled to −30° C. is added a solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[methoxy(methyl)-amino]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(129 mg, 0.25 mmole, from Example 52) keeping the temperature below −25°C. The reaction mixture is allowed to warm to 0° C., stir for 15 minsthen recooled to −30° C. and carefully quenched with ethyl acetate (2mL) and water (2 mL). The mixture is allowed to warm to room temperatureand partitioned between water and ether. The ether layer is dried oversodium sulfate, filtered and evaporated. The residue is dissolved in drytetrahydrofuran (5 mL), cooled to 0° C. and a solution of 1 Mphenylmagnesium bromide in tetrahydrofuran (0.75 mL, 0.75 mmole) addedvia syringe. The reaction mixture is stirred for 1 hour quenched withsaturated ammonium chloride and partitioned between water and ether. Theether layer is washed with water, brine, dried over sodium sulfate,filtered and the solid chromatographed on silica gel with 4:1hexane/ethyl acetate to give 22 mg of a white solid a mixture ofepimeric alcohols, MS (ES): m/z 536.2 (M+H).

EXAMPLE 75N,β,β-trimethyl-L-phenylalanyl-N-1-[(1S,2E,4R)-4-hydroxy-1-isopropyl-3-methyl-4-phenylbut-2-enyl}-N-1,3-dimethyl-L-valinamideand EXAMPLE 76N,β,β-trimethyl-L-phenylalanyl-N-1-[(1S,2E,4S)-4-hydroxy-1-isopropyl-3-methyl-4-phenylbut-2-enyl}-N-1,3-dimethyl-L-valinamide

The reaction described in Example 74 is repeated on twice the scale andthe mixture of alcohols are separated by hplc using acetonitrile/waterwith 0.02% formic acid from 5/95 to 100/0 over 1 hour. The first isomerisolated is arbitrarily assigned as the R alcohol, 45 mg of a whitesolid, as the formic acid salt which contains 7.2% of the S isomer, MS(ES): m/z 536.4 (M+H). The second isomer isolated is arbitrarilyassigned as the R alcohol, 83 mg of a white solid, as the formic acidsalt which contains 14.5% of the R isomer, MS (ES): m/z 536.4 (M+H).

EXAMPLE 77N,β,β-trimethyl-L-phenylalanyl-N-1-[(1S,2E)-4-hydroxy-1-isopropyl-3-methyl-4-(1,3-thiazol-2-yl)but-2-enyl]-N-1-,3-dimethyl-L-valinamide

To a slurry of lithium aluminum hydride (12 mg, 0.32 mmole) in ether (5mL) cooled to −30° C. is added a solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[methoxy(methyl)-amino]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(129 mg, 0.25 mmole, Example 52) keeping the temperature below −25° C.The reaction mixture is allowed to warm to 0° C., stir for 15 mins thenrecooled to −30° C. and carefully quenched with ethyl acetate (2 mL) andwater (2 mL). The mixture is allowed to warm to room temperature andpartitioned between water and ether. The ether layer is dried oversodium sulfate, filtered and evaporated. The residue is dissolved in drytetrahydrofuran (5 mL), cooled to 0° C. and a solution of2-lithiothiazole (prepared from thiazole (85 mg, 1.0 mmole) intetrahydrofuran (3 mL) at −78° C. and 2.5 M n-butyllithium (0.5 mL, 1.25mmole)) is added via syringe. The reaction mixture is stirred for 1 hourquenched with saturated ammonium chloride and partitioned between waterand ether. The ether layer is washed with water, brine, dried oversodium sulfate, filtered and the solid purified by HPLC to give 68 mg ofa white solid a mixture of epimeric alcohols, MS (ES): m/z 543.4 (M+H).

EXAMPLE 78N,β,β-trimethyl-L-phenylalanyl-N-1-[(1S,2E)-4-hydroxy-1-isopropyl-3-methylpent-2-enyl]-N-1,3-dimethyl-L-valinamide

To a slurry of lithium aluminum hydride (12 mg, 0.32 mmole) in ether (5mL) cooled to −30° C. is added a solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[methoxy(methyl)-amino]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(129 mg, 0.25 mmole, Example 52) keeping the temperature below −25°C.The reaction mixture is allowed to warm to 0° C., stir for 15 mins thenrecooled to −30° C. and carefully quenched with ethyl acetate (2 mL) andwater (2 mL). The mixture is allowed to warm to room temperature andpartitioned between water and ether. The ether layer is dried oversodium sulfate, filtered and evaporated. The residue is dissolved in drytetrahydrofuran (5 mL), cooled to 0° C. and a solution of 3.0M methylmagnesium bromide (0.25 mL, 0.75 mmole) is added via syringe. Thereaction mixture is stirred for 1 hour quenched with saturated ammoniumchloride and partitioned between water and ether. The ether layer iswashed with water, brine, dried over sodium sulfate, filtered and thesolid purified by HPLC to give 70 mg of a white solid a mixture ofepimeric alcohols, MS (ES): m/z 474.4 (M+H).

EXAMPLE 796-{[3,3-Dimethyl-2-(3-methyl-2-methylamino-3-phenyl-butyrylamino)-butyryl]-methyl-amino}-2,4,7-trimethyl-octa-2,4-dienoicacid

To a slurry of lithium aluminum hydride (48 mg, 1.28 mmole) in ether (20mL) cooled to −30° C. is added a solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[methoxy(methyl)-amino]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(516 mg, 1.0 mmole, Example 52) keeping the temperature below −25° C.The reaction mixture is allowed to warm to 0° C., stir for 15 min thenrecooled to −30° C. and carefully quenched with ethyl acetate (8 mL) andwater (8 mL). The mixture is allowed to warm to room temperature andpartitioned between water and ether. The ether layer is dried oversodium sulfate, filtered and evaporated. The residue is dissolved indichloromethane (10 mL), (carboethoxyehylidene)triphenylphosphorane(1.09 g, 3.0 mmole) is added. The reaction mixture is stirred for 1hour, then refluxed overnight. The reaction mixture is cooled to roomtemperature, evaporated to dyness and the residue purified bychromatography on silica gel eluting with 4:1 hexane/ethyl acetate togive 220 mg of a white solid. This solid 0.41 mmol) is treated withaqueous 1M lithium hydroxide solution (2.0 mL, 2.0 mmol), water (1.0 mL)and methanol (4.0 mL), to provide6-{[3,3-Dimethyl-2-(3-methyl-2-methylamino-3-phenyl-butyrylamino)-butyryl]-methyl-amino}-2,4,7-trimethyl-octa-2,4-dienoicacid (112 mg) as the corresponding trifluoroacetic acid salt as a whitesolid after preparative HPLC, MS (ES): m/z 514.4 (M+H).

EXAMPLE 80N,β,β-trimethyl-L-phenylalanyl-N-1-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-4-phenylbut-2-enyl]-N-1,3-dimethyl-L-valinamide

To a solution ofN,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[methoxy(methyl)-amino]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(129 mg, 0.25 mmole, Example 52) in tetrahydrofuran (5 mL) at 0° C. isadded 1M phenylmagnesium bromide in tetrahydrofuran (0.75 mL, 0.75mmole) via syringe. The reaction mixture is stirred for 1 hour quenchedwith saturated ammonium chloride and partitioned between water andether. The ether layer is washed with water, brine, dried over sodiumsulfate, filtered and the residue purified by HPLC to give 36 mg of awhite solid, MS (ES): m/z 534.4 (M+H).

EXAMPLE 81(2S)-N-[(1S,2E)-4-hydroxy-1-isopropyl-3-methyl-2-butenyl]-N,3,3-trimethyl-2-{{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamide

ToN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-3-carboxy-1-isopropylbut-2-enyl]-N¹,3-dimethyl-L-valinamide(200 mg) in acetonitrile (20 mL) at 25° C. is added1-hydroxybenzotriazole hydrate (68 mg) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (108 mg).After 2 hours sodium borohydride (92 mg) is added. After 18 hours, tothe heterogeneous reaction mixture is added water followed by 2N aqueoushydrochloric acid. Concentration in vacuo and purification by reversephase HPLC (0.01% aqueous trifluoroacetic acid/acetonitrile gradientsystem) gave the trifluoracetic acid salt of the title compound as awhite powder (148 mg). MS (ES): m/z 460.4 (M+H).

EXAMPLE 82N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamideEXAMPLE 834-(dimethylsulfonio)-N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamidetrifluoroacetic acid

Following the procedure described in J. Org. Chem. 1994, 59(7), 1771, toa solution of oxalyl chloride (0.17 mmol) in dichloromethane at −78° C.is added a solution of dimethylsulfoxide (0.348 mmol, 0.025 mL) indichloromethane dropwise. After 10 min a solution of(2S)-N-[(1S,2E)-4-hydroxy-1-isopropyl-3-methyl-2-butenyl]-N,3,3-trimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamidetrifluoroacetic acid (83 mg, from Example 81) in dichloromethane isadded dropwise. After 20 min triethylamine (0.725 mmol, 0.1 mL) is addeddropwise. The reaction mixture is allowed to warm to 25° C. then washedwith water and dried over anhydrous sodium sulfate. Concentration invacuo and purification by reverse phase HPLC (0.01% aqueoustrifluoroacetic acid/acetonitrile gradient system) gave thetrifluoracetic acid salt ofN,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamideas a white powder (43 mg) MS (ES): m/z 458.1 (M+H) and4-(dimethylsulfonio)-N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N¹,3-dimethyl-L-valinamidetrifluoroacetate as a clear glass (28 mg) MS (ES): m/z 518.0 (M⁺).

EXAMPLE 84N,β,β-trimethyl-L-phenylalanyl-N¹-[(1S,2E)-1-isopropyl-3-methyl-4-oxo-2-pentenyl]-N¹,3-dimethyl-L-valinamide

According to the procedures in J. Am. Chem. Soc., 1980, 3270 andTetrahedron Letters, 1984, 5733 a solution of ethyl(E,4S)-4-[((2S)-3,3-dimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(250 mg) in tetrahydrofuran (5 mL) at −40° C. under nitrogen atmosphereis treated with Tebbe reagent (0.5 M in toluene, 0.55 mL, 1.1 mmol)dropwise over 1 min. After 15 min the reaction mixture is allowed towarm to 25° C. After 2 h 15% aqueous sodium hydroxide (0.15 mL) is addedto the red solution at −10° C. The reaction mixture is allowed to warmto 25° C. After 10 min gas evolution ceased and the green mixture isdiluted with ether, dried over anhydrous sodium sulfate and filteredthrough diatomaceous earch. Concentration in vacuo gave a yellow oil(385 mg) (MS m+h=500). A portion of this material (285 mg) is dissolvedin tetrahydrofuran (8 mL) and treated with 1N aqueous hydrochloric acid(0.4 mL). After 18 h the mixture is filtered through diatomaceous earthand purified by reverse phase HPLC (0.01% aqueous trifluoroaceticacid/acetonitrile gradient system) to give the trifuoroacetic acid saltof the title compound as a pale yellow powder. MS (ES): m/z 472.6 (M+H)

EXAMPLE 85N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S)-1-[(Z)-(2,5-dioxo-4-imidazolidinylidene)methyl]-2-methylpropyl}-N¹,3-dimethyl-L-valinamide

To a solution of diethyl 2,4-dioxoimidazolidine-5-phosphonate (N. A.Meanwell et. al. J. Org. Chem. 1991, 56(24), 6897) (192 mg, 0.81 mmol)in ethanol (2 mL) is added a solution of lithium hydroxide hydrate (31mg, 0.73 mmol) in water (0.2 mL). The resulting solution is transferred,using a minimum of ethanol, to a solution ofN-{1-[(1-Formyl-2-methyl-propyl)-methyl-carbamoyl]-2,2-dimethyl-propyl}-3-methyl-2-methylamino-3-phenyl-butyramide(114 mg, 0.27 mmol, Reference Example 12) in ethanol (0.5 mL). After 3days the reaction mixture is concentrated in vacuo and purified byreverse phase HPLC (0.01% aqueous trifluoroacetic acid/acetonitrilegradient system) to give the trifluoroactic acid salt of the titlecompound (3:1 mixture with its double bond isomer) as a white powder (17mg).

HRMS (ESI) calcd for C₂₇H₄₁N₅O₄ 499.31602, found 500.32152 [M+H]

REFERENCE EXAMPLE 13 tert-butyl(1S,2E)-3-cyano-1-isopropyl-2-propenyl(methyl)carbamate

To a solution of tert-butyl(1S)-1-formyl-2-methylpropyl(methyl)carbamate (WO 99/32509) (500 mg, 202mmol) in dicloromethane (5 mL) is added(triphenylphosphoranylidene)acetonitrile (700 mg, 2.32 mmol). After 1 hadditional (triphenylphosphoranylidene)acetonitrile is added (350 mg,1.16 mmol). After 24 h the reaction mixture is concentrated in vacuo andchromatographed (silica gel, hexane/ether) to give the title compound asan oil (410 mg). Analysis for C13H22N2O2: Calc'd: C, 65.52; H, 9.3; N11.75. Found: C, 65.3; H, 9.05; N,11.51. MS (ES): m/z 239.1 (M+H)

REFERENCE EXAMPLE 14 (E,4S)-5-methyl-4-(methylamino)-2-hexenenitrile

To a solution of tert-butyl(1S,2E)-3-cyano-1-isopropyl-2-propenyl(methyl)carbamate (400 mg, 1.68mmol, from Reference Example 13) in dichloromethane (5 mL) is addedtrifluoroacetic acid (2 mL). After 2 h the reaction mixture isconcentrated in vacuo. Dissolution in dichloromethane followed byconcentration in vacuo (3×) gave the trifluoroacetic acid salt of thetitle compound as an oil. MS (ES): m/z 139.1 (M+H)

REFERENCE EXAMPLE 15 tert-butyl(1S)-1-{[[(1S,2E)-3-cyano-1-isopropyl-2-propenyl](methyl)amino]carbonyl}-2,2-dimethylpropylcarbamate

To a solution of (E,4S)-5-methyl-4-(methylamino)-2-hexenenitriletrifluoroacetic acid (−1.68 mmol, from Reference Example 14),2-tert-Butoxycarbonylamino-3,3-dimethyl-butyric acid (389 mg, 1.68mmol), (benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate (PyBOP, 916 mg, 1.76 mmol) in dichloromethane (5 mL)at OC is added Hunig's base (0.877 mL, 5.04 mmol). After 5 min thecooling bath is removed. After 24 h 5% aqueous citric acid is added. Theaqueous phase is washed with dichloromethane three times. The combinedorganic extracts are dried over anhydrous magnesium sulfate andconcentrated in vacuo to give an oil. Chromatography (silica gel,hexane/ether) gave the title compound as a yellow foam (200 mg). MS(ES): m/z 352.0 (M+H)

REFERENCE EXAMPLE 16(2S)-2-amino-N-[(1S,2E)-3-cyano-1-isopropyl-2-propenyl]-N,3,3-trimethylbutanamide

To a solution of tert-butyl(1S)-1-{[[(1S,2E)-3-cyano-1-isopropyl-2-propenyl](methyl)amino]carbonyl}-2,2-dimethylpropylcarbamate(180 mg, from Reference Example 15) in dichloromethane (5 mL) is addedtrifluoroacetic acid (2 mL). After 20 min the reaction mixture isconcentrated in vacuo. Dissolution in dichloromethane followed byconcentration in vacuo (3×) gave the trifluoroacetic acid salt of thetitle compound as a yellow foam (210 mg). MS (ES): m/z 252.2 (M+H)

REFERENCE EXAMPLE 17 tert-butyl(1S)-1-{[((1S)-1-{[[(1S,2E)-3-cyano-1-isopropyl-2-propenyl](methyl)amino]carbonyl}-2,2-dimethylpropyl)amino]carbonyl}-2-methyl-2-phenylpropyl(methyl)carbamate

To(2S)-2-amino-N-[(1S,2E)-3-cyano-1-isopropyl-2-propenyl]-N,3,3-trimethylbutanamidetrifluoroacetic acid (179 mg, from Reference Example 16),O-(7-azabenzotriazol-1-yl)-N,N, N′,N′-tetramethyluroniumhexafluorophosphate (HATU, 297 mg) and 1-Hydroxy-7-azabenzotriazole(HOAt, 6.8 mg) is added diisopropylethylamine (252 uL) followed bydichloromethane (9 mL) and(2S)-2-[(tert-butoxycarbonyl)(methyl)amino]-3-methyl-3-phenylbutanoicacid (162 mg). After 18 h 5% aqueous citric acid is added. The aqueousphase is washed with dichloromethane three times. The combined organicextracts are dried over anhydrous magnesium sulfate and concentrated invacuo to give a yellow oil. Chromatography (silica gel, hexane/ether)gave the title compound as a yellow foam (176 mg). MS (ES): m/z 541.1(M+H)

EXAMPLE 86(2S)-N-[(1S,2E)-1-isopropyl-3-(1H-tetraazol-5-yl)-2-propenyl]-N,3,3-trimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamide

Following the procedure outlined in J. Org. Chem., 1993, 58, 4139,tert-butyl(1S)-1-{[((1S)-1-{[[(1S,2E)-3-cyano-1-isopropyl-2-propenyl](methyl)amino]carbonyl}-2,2-dimethylpropyl)amino]carbonyl}-2-methyl-2-phenylpropyl(methyl)carbamate(40 mg, from Reference Example 17), azidotrimethylsilane (20 uL, 17 mg)and dibutyltin oxide (2 mg) in toluene (1.6 mL) is heated at 100° C. for18 h. Additional azidotrimethylsilane (10 uL) and dibutyltin oxide (0.5mg) is added and the reaction mixture heated further. The reactionmixture is concentrated in vacuo. Dissolution in methanol followed byconcentration in vacuo (3×) gave an oil. Chromatography (silica gel,dichloromethane/methanol) gave an oil (20 mg). A solution of thismaterial in dichloromethane (2 mL) is treated with trifluoroacetic acid(1 mL). After 1 h the reaction mixture is concentrated in vacuo.Dissolution in dichloromethane followed by concentration in vacuo (3×)gave the trifluoroacetic acid salt of the title compound as a pink solid(14 mg). MS (ES): m/z 484.0 (M+H)

REFERENCE EXAMPLE 18 tert-butylmethyl[(1S)-2-methyl-1-(4-morpholinylcarbonyl)-2-phenylpropyl]carbamate

To a cooled (0°C., ice bath) solution of(2S)-2-[(tert-butoxycarbonyl)(methyl)amino]-3-methyl-3-phenylbutanoicacid (1.0 g, 3.30 mmol, WO 99/32509) andbenzotriazole-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate(1.46 g, 3.30 mmol) in anhydrous dichloromethane (5 ml) is addeddiisopropylethylamine (1.15 ml, 6.60 mmol) under an inert atmosphere. Tothis solution is added a solution of morpholine (0.313 ml, 3.60 mmol) indichloromethane (2.5 ml). After stirring at 0° C. for 5-10 minutes, thecooling bath is removed, and the resulting reaction mixture is stirredat room temperature for 15-20 hours. The mixture is diluted with water,and the aqueous layer is extracted with ethyl acetate (3 times). Thecombined extracts are washed with saturated sodium chloride solution,dried over sodium sulfate, filtered and concentrated in vacuo. Theresidue is chromatographed (silica gel, flash column), to give a darkyellow oil. (0.943 g, 76%) MS [M+H] 377.2

REFERENCE EXAMPLE 19 tert-butyl(1S)-1-formyl-2-methyl-2-phenylpropyl(methyl)carbamate

To a cooled solution (0° C., ice bath) of tert-butylmethyl[(1S)-2-methyl-1-(4-morpholinylcarbonyl)-2-phenylpropyl]carbamate(0.870 g, 2.32 mmol, from Reference Example 18) in anhydroustetrahydrofuran (3.0 ml), 1.0M lithium aluminum hydride (5.8 ml, 5.8mmol) in tetrahydrofuran is added dropwise over 30 minutes. The reactionmixture stirred at room temperature for 30 minutes and is quenched with5% potassium hydrogen sulfate and stirred at room temperature for 15minutes. The mixture is washed with ether (3 times). The organic layeris then collected and dried with sodium sulfate and concentrated invacuo to give a colorless oil. The crude product is used without furtherpurification. (0.940 g, 80%) MS [M+H] 292.2

REFERENCE EXAMPLE 20ethyl(2E,4S)-4-[(N-{(2S)-2-[(tert-butoxycarbonyl)(methyl)amino]-3-methyl-3-phenylbutyl}-3-methyl-L-valyl)(methyl)amino]-2,5-dimethyl-2-hexenoate

To a stirred slurry of tert-butyl(1S)-1-formyl-2-methyl-2-phenylpropyl(methyl) carbamate (104 mg, 0.40mmole, Reference Example 19) with ethyl4-[(2-Amino-3,3-dimethyl-butyryl)-methyl-amino]-2,5-dimethyl-hex-2-enoicacid ethyl ester (122 mg, 0.40 mmole) in MeOH (2.5 ml), a solution ofzinc chloride (32.7 mg, 0.6 eq) and sodium cyanoborohydride (34.4 mg,0.48 mmol) in MeOH (2.5 ml) is added over 5 minutes at room temperature.The resulting mixture is stirred for 15-24 hours at room temperatureunder an inert atmosphere. After concentration, the residue is taken upin ethylacetate, washed with sodium bicarbonate. The aqueous phase isextracted with ethylacetate. The combined organic layers is washed withbrine, dried and is chromatographed (silica gel, flash column) to give awhite solid. (0.164 mg, 81%) M.W. 587.8 [M+H] 588.3

REFERENCE EXAMPLE 21(2E,4S)-4-[(N-{(2S)-2-[(tert-butoxycarbonyl)(methyl)amino]-3-methyl-3-phenylbutyl}-3-methyl-L-valyl)(methyl)amino]-2,5-dimethyl-2-hexenoicacid

Ethyl(2E,4S)-4-[(N-{(2S)-2-[(tert-butoxycarbonyl)(methyl)amino]-3-methyl-3-phenylbutyl}-3-methyl-L-valyl)(methyl)amino]-2,5-dimethyl-2-hexenoate (50 mg,0.085 mmol, Reference Example 20) is dissolved in methanol (1 ml),tetrahydrofuran (1 ml). To this solution is added water (0.5 ml) andsolid lithium hydroxide (4.1 mg). The resulting mixture is stirred atroom temperature for 15 hours. The organic solvents are removed invacuo, and the residual aqueous mixture is cooled with an ice waterbath, and acidified to pH 5.5-6.0 with aqueous 1 N aqueous citric acidsolution. The precipitate is collect by filtration, and the solid iswashed with a cold water, and dried over high vacuum. Alternatively, theproduct can be purified by using preparative HPLC. (40 mg, 80%) MS [M+H]588.2

EXAMPLE 87(2E,4S)-2,5-dimethyl-4-(methyl{3-methyl-N-[(2S)-3-methyl-2-(methylamino)-3-phenylbutyl]-L-valyl}amino)-2-hexenoicacid

(2E,4S)-4-[(N-{(2S)-2-[(tert-butoxycarbonyl)(methyl)amino]-3-methyl-3-phenylbutyl}-3-methyl-L-valyl)(methyl)amino]-2,5-dimethyl-2-hexenoicacid (40 mg, 0.068 mmol, from reference example 21) is dissolved indichloromethane (3 ml) and treated with 4N hydrogen chloride (0.068 ml,0.27 mmol) at room temperature for 30 minutes. The reaction mixture isthen concentrated in vacuo and triturated with ether to give a whitesolid (29 mg, 87%). The crude material is purified by reverse phaseHPLC. MS [M+H] 488.62

EXAMPLE 88(2S)-N-[(1S,2E)-4-(1H-imidazol-2-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]--trimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamide

To a solution of ethyl(E,4S)-4-[tert-butoxycarbonyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(2.8 g, 9.35 mmole) in 43 ml anhydrous tetrahydrofuran is added 1 Maqueous lithium hydroxide (46.7 ml, 46.76 mmole), methanol (90.5 mL) andwater (23.8 ml). The reaction mixture is stirred at room temperatureovernight. The solvent is removed in vacuo. The residue is partitionbetween ethyl acetate and water. The aqueous layer is acidified with 1 Nhydrochloric acid dropwise in ice bath until pH 4.5. Organic layer isseparated. The aqueous layer is extracted twice with ethyl acetate. Thecombined organic extracts are washed with brine, dried over sodiumsulfate, filtered, concentrated in vacuo to give 2.42 g (95%) of(2E,4S)-4-[(tert-butoxycarbonyl)(methyl)amino]-2,5-dimethylhex-2-enoicacid as a white solid. MS(ES⁻): (M−H)=270.2

A solution of this carboxylic acid (1.56 g, 5.75 mmole) in 11.61 mltetrahydrofuran is treated with N,O-dimethylhydroxylamine hydrochloride(954 mg, 9.8 mmol) which is dissolved in 11.61 ml water. The pH isadjusted to 4.5 with 1 N sodium hydroxide solution.1-[3-(dimethylamino)propyl]-3-ethylcarbodiimidehydrochloride (2.76 g,14.4 mmol) in water (39.4 ml) is added dropwise over a period of 25minutes to the solution of carboxylic acid andN,O-dimethylhydroxylamine, while the pH is maintained to a constantvalue of 4.5. After 2.5 hours, the solution is saturated with sodiumchloride and extracted with ethyl acetate. The organic extract is driedover sodium sulfate, filtered, concentrated in vacuo to give(1S,2E)-1-isopropyl-4-[methoxy(methyl)amino]-3-methyl-4-oxobut-2-enyl(methyl)carbamate903.9 mg (50%) as a light yellow oil. MS(ES⁺): (M+Na)=337.1

To a solution of imidazole (10.0 g, 146.9 mmole) in 100 ml dimethylsulfoxide at room temperature under nitrogen is added sodium hydride(6.46 g, 161.6 mmole). The solution is stirred at 80° C. for 2 hours.Chloromethyl ethyl ether (16.35 ml, 176.3 mmole) is added dropwise. Thereaction mixture is stirred for another 2 hour at 80° C., cooled to roomtemperature, poured into 5% aqueous solution and extracted withdichloromethane (4×200 ml). The combined organic extracts are dried oversodium sulfate, filtered, concentrated in vacuo. The residue is freshchromatographed in hexane/ethyl acetate from 5:1 to 2:1 to give1-ethoxymethyl-1H-imidazole as a brown oil 1.68 g (90%) MS(ES⁺):(M+H)=127.0 To the solution of 1-ethoxymethyl-1H-imidazole (1.24 g, 0.98mmole) in 3.92 ml anhydrous tetrahydrofuran at −78° C. is addedn-butyllithium (1.6 M in hexane) over 5 minutes. Reaction mixture isstirred at −78° C. for 1.5 hour, then(1S,2E)-1-isopropyl-4-[methoxy(methyl)amino]-3-methyl-4-oxobut-2-enyl(methyl)carbamate(427 mg, 1.36 mmole) in 2 ml anhydrous tetrahydrofuran is added over 10minutes. The reaction mixture is stirred at −78° C. for 1 hour. Then, itis warmed to room temperature, stirred for 1 hour at room temperature.The reaction mixture is diluted with dichloromethane/water. The organiclayer is washed with 5% sodium bicarbonate, dried over sodium sulfate,filtered, concentrated in vacuo. The residue is fresh chromatographed inhexane/ethyl acetate from 3:1 to 1:1 to give light yellow oil:[4-(1-Ethoxymethyl-1H-imidazol-2-yl)-1-isopropyl-3-methyl-4-oxo-but-2-enyl]-methyl-carbamicacid tert-butyl ester (120 mg, 23%) MS(ES⁺): (M+H)=380.2

[4-(1-Ethoxymethyl-1H-imidazol-2-yl)-1-isopropyl-3-methyl-4-oxo-but-2-enyl]-methyl-carbamicacid tert-butyl ester (496 mg, 1.31 mmole) is stirred in 4N hydrochloricacid/dioxane at room temperature under nitrogen for 2 hours. The solventis removed in vacuo. Dichloromethane is added to the residue andevaporated (this step is repeated twice) to give a white solid aminointermediate which then stirred with2-tert-Butoxycarbonylamino-3,3-dimethyl-butyric acid (410.5 mg, 1.78mmole), 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT,531 mg, 1.77 mmole) and triethylamine (0.49 ml, 3.55 mmole) in 4 mldichloromethane at room temperature overnight. The reaction mixture isdiluted with dichloromethane/water. Organic layer is separated. Theaqueous layer is extracted twice with dichloromethane. The combinedorganic extracted are washed with citric acid (4 ml), sodium carbonate(6 ml) and brine, dried over sodium sulfate, filtered, concentrated invacuo. The residue is flash chromatographed with hexane/ethyl acetatefrom 5:1 to 1:1 to give a white crystals,(1-{[4-(-Ethoxymethyl-1H-imidazol-2-yl)-1-isopropyl-3-methyl-4-oxo-but-2-enyl]-methyl-carbonyl}-2,2-dimethyl-propyl)-carbamicacid tert-butyl ester: 171 mg (26%). MS(ES⁺): (M+H)=493.2

Following above procedure,(1-{[4-(-Ethoxymethyl-1H-imidazol-2-yl)-1-isopropyl-3-methyl-4-oxo-but-2-enyl]-methyl-carbonyl}-2,2-dimethyl-propyl)-carbamicacid tert-butyl ester (171 mg, 0.34 mmole) is coupled with(2S)-2-(tert-Butoxycarbonyl-methyl-amino)-3-phenyl-3-methyl-butyric acid(128 mg, 0.42 mmole) in the presence of3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT, 104 mg,0.35 mmole) and triethylamine (1.4 ml, 0.69 mmole) and 2.5 mldichloromethane. The product isolated by reverse phase HPLC (0.01%trifluoroacetic acid in water/acetonitrile) to give a yellow oil,[1-(1-{[4-(-Ethoxymethyl-1H-imidazol-2-yl)-1-isopropyl-3-methyl-4-oxo-but-2-enyl]-methyl-carbamoyl}-2,2-dimethyl-propylcarbamoyl)-2-methyl-2-phenyl-propyl]-methyl-carbamicacid tert-butyl ester: 46 mg (20%) MS(ES⁺): (M+H)=682.4

[1-(1-{[4-(-Ethoxymethyl-1H-imidazol-2-yl)-1-isopropyl-3-methyl-4-oxo-but-2-enyl]-methyl-carbamoyl}-2,2-dimethyl-propylcarbamoyl)-2-methyl-2-phenyl-propyl]-methyl-carbamicacid tert-butyl ester (45 mg, 0.066 mmole) is treated with 2 mltrifluoroacetic acid in 2 ml dichloromethane at room temperature for 2hours. The solvent is evaporated and pumped in oven overnight to give 8mg white solid product,(2S)-N-[(1S,2E)-4-(1H-imidazol-2-yl)-1-isopropyl-3-methyl-4-oxobut-2-enyl]-N,3,3-trimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamide:8 mg (16%). MS(ES⁺): (M+H)=524.4

EXAMPLE 89N,β,β-trimethyl-L-phenylalanyl-N¹,3-dimethyl-N¹-{(1S)-2-methyl-1-[(Z)-(3-methyl-2,5-dioxoimidazolidin-4-ylidene)methyl]propyl}-L-valinamide

A mixture of 1-methylhydantoin (20.25 g, 180 mmole) and acetic acid (100ml) is heated to 90° C. Bromine (10 ml, 195 mmole) is introduceddropwise at such a rate that instant decolorization occurred. After theaddition is completed, the reaction mixture is stirred at 90° C. for 60minute and at room temperature overnight. The acetic acid is decantedfrom white precipitate and removed in vacuo. The residue is combinedwith the precipitate and suspended in diethylether (200 ml).Triethylphosphite (32 ml, 180 mmole) is added portionwise with stirringand cooling. A solution resulted which on continued stirring, yieldtrace of white precipitate. The mixture is diluted with diethyl etherand allowed to stand overnight. Decanted the solvent, and pumped invacuum oven to give white crystal ylide,(3-Methyl-2,5-dioxo-imidazolidin-4-yl)-phosphonic acid diethyl ester:16.57 g (36.8%) MS(ES⁺): (M+H)=251.0

The yield, (3-methyl-2,5-dioxo-imidazolidin-4-yl)-phosphonic aciddiethyl ester (0.915 g, 3.0 mmol) in ethanol (9 mL) is added to lithiumhydroxide monohydrate (139 mg, 2.1 mmol) in 0.9 ml water to give ayellow solution. It is then added toN-{1-[(1-formyl-2-methyl-propyl)-methyl-carbamoyl]-2,2-dimethyl-propyl}-3-methyl-2-methylamino-3-phenyl-butyramide(˜1 mmol, from Reference Example 12) in 3 mL ethanol. The reactionmixture is stirred at room temperature for 60 hours. Solvent isevaporated. The residue is diluted with dichloromethane/water. Theaqueous layer is extracted twice with dichloromethane. The combinedorganic extracts are washed with brine, dried oversodium sulfate,filtered and concentrated in vacuo. The residue is chromatographed byreverse phase HPLC (0.01% trifluoroacetic acid in water/acetonitrile) togive the product as a white solid,N,beta,beta-trimethyl-L-phenylalanyl-N¹,3-dimethyl-N¹-{(1S)-2-methyl-1-[(Z)-(3-methyl-2,5-dioxoimidazolidin-4-ylidene)methyl]propyl}-L-valinamide:25mg (4%) MS(ES⁺): (M+H)=514.3

EXAMPLE 90N-{1-[(5-Hydroxy-1-isopropyl-3-methyl-4-oxo-pent-2-enyl)-methyl-carbamoyl]-2,2-dimethyl-propyl}-3-methyl-2-methylamino-3-phenyl-butyramide

To a solution oftrimethyl-(2-tributylstannanylmethoxymethoxy-ethyl)-silane (933 mg, 2.07mmole, E. Fernandez-Megia and S. V. Ley; Synlett 2000, 4, 455-8) in 6 mLtetrahydrofuran at −78° C. is added dropwise 1.6 M n-butyllithiumsolution in hexanes (1.25 mL, 2.0 mmole). The resulting yellow solutionis stirred for 10 mins at −78° C. Then a solution ofN,β,β-trimethyl-L-phenylalanyl-N¹{(1S,2E)-1-isopropyl-4-[methoxy(methyl)amino]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide(258 mg, 0.5 mmole, Example 52) in tetrahydrofuan (5 mL) at −78° C. isadded dropwise and the reaction mixture stirred at −78° C. for 15 minsand then at −60° C. for 45 mins. The reaction is quenched with asaturated aqueous solution of ammonium chloride (2 mL), warmed to roomtemperature and partioned between ethyl acetate (50 mL) and saturatedaqueous solution of ammonium chloride (50 mL). The aqueous layer isextracted with ethyl acetate (30 mL) and the combined organic layer iswashed with water, brine, dried over sodium sulfate, filtered and theresidue treated with a mixture of trifluoroacetic acid (10 mL) anddichloromethane (5 mL) at 0° C. and stirred for 1 hour, evaporated andthen purified by HPLC to give 52 mg of an off-white solid, MS (ES): m/z488.4 (M+H).

REFERENCE EXAMPLE 22 3,3-dimethyl-3-phenyl propionic acid

Following a literature procedure, (Leffler, J. E.; Barbas, J. T. JACS.1981, 103(26), 7768-7773), to a mixture of magnesium turnings (9.5 g)and neophyl chloride (7.5 mL) in tetrahydrofuran (100 mL) is added aspatula tip of iodine and 1,2-dibromoethane (1 mL). The mixture isheated gently with under a heat gun to initiate a vigorous reaction. Asolution of neophyl chloride (57 mL) in tetrahydrofuran (360 mL) isadded dropwise over the course of 90 minutes. Following completion ofthe addition, the reaction mixture is stirred for an additional fivehours, then solid carbon dioxide (25 g) is added to the flask. After 10minutes, the flask is transferred to a 40° C. water bath until allcarbon dioxide had sublimed. The solvent is then removed under reducedpressure to afford the title compound (58 g, 81%) as a white paste. MS(ES⁺): (M+H)=178.8

REFERENCE EXAMPLE 23(4S)-4-isopropyl-3-(3-methyl-3-phenylbutanoyl)-1,3-oxazolidin-2-one

To a solution of 3,3-dimethyl-3-phenyl propionic acid (2.1 g, 12 mmol,from Reference Example 22) in anhydrous tetrahydrofuran (24 mL) is addedunder inert atmosphere triethylamine (2.0 mL) and the mixture is cooledto −78° C. in a dry-ice acetone bath. Pivaloyl chloride (1.5 mL, 13mmol) is added dropwise, causing the immediate formation of a whiteprecipitate. The reaction mixture is allowed to sit for 15 minutes at−78° C. and is then stirred at 0° C. in an ice-water bath. In a separateflask, a solution of (S)-4-isopropyl-2-oxazolidinone (1.5 g, 12 mmol) inanhydrous tetrahydrofuran (27 mL) is prepared under inert atmosphere andcooled to −35° C. in a dry-ice/acetone bath. A small amount oftriphenylmethane (<5 mg) is added as an indicator of deprotonation.n-Butyllithium (1.6 M solution in hexanes, 7.7 mL, 12 mmol) is addeddropwise via syringe. After 30 minutes of stirring at 0° C., the flaskcontaining the mixed anhydride is re-cooled to −78° C. in adry-ice/acetone bath. The solution of the lithium anion of theoxazolidinone is added to the mixed anhydride solution via cannula. Thesource flask is washed twice with tetrahydrofuran (4 mL×2) and thesewashings are also transferred via cannula to the mixed anhydridesolution. The reaction mixture is stirred at −78° C. for 1 hour, 0° C.for 1 hour, and allowed to warm to room temperature overnight. Thefollowing morning, water (˜15 mL) is added and stirring is continued for20 minutes. The aqueous phase is extracted thrice with diethyl ether.The combined extracts are washed with saturated aqueous sodium chloride,dried over sodium sulfate, decanted, and concentrated under reducedpressure to afford (3.6 g,>100% crude) of a pale yellow oil. The crudematerial is purified by flash chromatography (ethyl acetate/hexanes) toafford 1.5 g (44%) of a clear, colorless oil. MS (ES⁺): (M+H)=290.0

REFERENCE EXAMPLE 24(4S)-3-[(2S)-2,3-dimethyl-3-phenylbutanoyl]-4-isopropyl-1,3-oxazolidin-2-one

A solution of(4S)-4-isopropyl-3-(3-methyl-3-phenylbutanoyl)-1,3-oxazolidin-2-one(0.23 g, 0.79 mmol, from Reference Example 23) in anhydroustetrahydrofuran (3 mL) is cooled to −78° C. in a dry-ice/acetone bathwhile stirring under a nitrogen atmosphere. Sodium hexamethylsilazide(1.0 M solution in tetrahydrofuran, 0.95 mL, 0.95 mmol) is addeddropwise to the solution via syringe. After stirring for 30 minutes at−78° C., iodomethane (0.25 mL, 4.0 mmol) in tetrahydrofuran (1 mL) isadded dropwise. The reaction mixture is stirred overnight (18 hours)while warming to room temperature. The reaction mixture is diluted withwater (10 mL) and ethyl acetate (10 mL). The aqueous phase is extractedthrice with ethyl acetate. The combined extracts are washed withsaturated aqueous sodium hydrogen carbonate and saturated aqueous sodiumchloride, dried over sodium sulfate, decanted, and concentrated underreduced pressure to afford a clear yellow oil, which slowly solidifiedto an amorphous solid under high vacuum. The crude material is purifiedby flash chromatography (ethyl acetate/hexanes) to afford a crystallinesolid (0.17 g, 71%). MS (ES⁺): (M+H)=303.9

REFERENCE EXAMPLE 25 (2S)-2,3-dimethyl-3-phenylbutanoic acid

To a 0° C. solution of(4S)-3-[(2S)-2,3-dimethyl-3-phenylbutanoyl]-4-isopropyl-1,3-oxazolidin-2-one(0.13 g, 0.43 mmol, from Reference Example 24) in tetrahydrofuran (6 mL)and water (2 mL) is added 30% aqueous hydrogen peroxide (2.4 mL). Afterone to two minutes of stirring, lithium hydroxide monohydrate (36 mg) isadded and the reaction mixture is allowed to stir overnight whilewarming to room temperature. On the following day, the reaction mixtureis cooled to 0° C. and aqueous sodium sulfite solution (1.5 M, 1.3 mL)is added. After stirring for an hour at room temperature, the reactionmixture is concentrated under reduced pressure. The aqueous residue isextracted twice with dichloromethane, then acidified to pH 1 with 10%aqueous hydrochloric acid solution. The acidified aqueous phase is thenextracted thrice with ethyl acetate. The combined organic extracts arewashed with saturated aqueous sodium chloride, dried over anhydroussodium sulfate, decanted, and concentrated under reduced pressure togive a white crystalline solid (60 mg, 72%). Mp 69-72° C. MS (ES⁻):(M−H)=191.0

EXAMPLE 91 Ethyl(E,4S)-4-[((2S)-2-{[(2S)-2,3-dimethyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate

To a cooled (0° C., ice bath) solution of(2S)-2,3-dimethyl-3-phenylbutanoic acid (0.11 g, 0.57 mmol, fromReference Example 25), hydroxybenzotriazole (93 mg, 0.69 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimine hydrochloride (0.15 g,0.80 mmol) in anhydous dimethylformamide (2 ml) is addedN-methylmorpholine (95 uL, 0.86 mmol) via syringe under an inertatmosphere. After stirring for 15 minutes at 0° C., the cooling bath isremoved, and the resulting mixture is stirred for 1 hour. The solutionis cooled at 0° C. (ice water bath), and to this mixture is added asolution of ethyl(2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoatehydrochloride (0.56 mmol) in anhydrous dimethylformamide (2 ml). Thecooling bath is removed, and the resulting mixture is stirred for 22hours at room temperature under an inert atmosphere. The mixture isdiluted with water, and the aqueous layer is extracted with ethylacetate (3 times). The combined extracts are washed with saturatedaqueous sodium chloride, dried over anhydrous sodium sulfate, filteredand concentrated in vacuo. The residue is chromatographed (silica gel,flash column), to provide a white foam (0.20 g, 71%). MS (ES⁺):(M+H)=487.0

EXAMPLE 92(E,4S)-4-[((2S)-2-{[(2S)-2,3-dimethyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoicacid

Ethyl(E,4S)-4-[((2S)-2-{[(2S)-2,3-dimethyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(0.17 g, 0.35 mmol, from Example 91) is dissolved in methanol (1.2 mL)and tetrahydrofuran (1.2 mL) and cooled to 0° C. (ice water bath). Tothis solution is added water (0.6 mL) and aqueous lithium hydroxidemonohydrate (30 mg, 0.70 mmol). The cooling bath is removed, and theresulting mixture is stirred at 50° C. for 15 hours. Solvents areremoved in vacuo to provide a beige foam, which is then dissolved inwater. The solution is acidfied to pH 1 with 10% aqueous hydrochloricacid solution and then extracted thrice with ethyl acetate. The combinedextracts are washed with saturated sodium chloride solution, dried overanhydrous sodium sulfate, decanted, and concentrated under reducedpressure to afford a white solid (0.16 g, 100%). MS (ES⁺): (M+H)=459.0

REFERENCE EXAMPLE 26(4S)-4-benzyl-3-(3-methyl-3-phenylbutanoyl)-1,3-oxazolidin-2-one

To a solution of 3,3-dimethyl-3-phenyl propionic acid (10 g, 57 mmol,from Reference Example 22) in anhydrous tetrahydrofuran (120 mL) isadded under inert atmosphere triethylamine (10 mL) and the mixture iscooled to −78° C. in a dry-ice acetone bath. Pivaloyl chloride (7.4 mL,60 mmol) is added dropwise, causing the immediate formation of a whiteprecipitate. The reaction mixture is allowed to sit for 15 minutes at−78° C. and is then stirred at 0° C. in an ice-water bath. In a separateflask, a solution of (S)-4-benzyl-2-oxazolidinone (9.9 g, 56 mmol) inanhydrous tetrahydrofuran (130 mL) is prepared under inert atmosphereand cooled to −35° C. in a dry-ice/acetone bath. A small amount oftriphenylmethane (<5 mg) is added as an indicator of deprotonation.n-Butyllithium (1.6 M solution in hexanes, 40 mL, 64 mmol) is addeddropwise via syringe. After 30 minutes of stirring at 0° C., the flaskcontaining the mixed anhydride is re-cooled to −78° C. in adry-ice/acetone bath. The solution of the lithium anion of theoxazolidinone is added to the mixed anhydride solution via cannula. Thesource flask is washed twice with tetrahydrofuran (4 mL×2) and thesewashings are also transferred via cannula to the mixed anhydridesolution. The reaction mixture is stirred at −78° C. for 1 hour and at0° C. for 1 hour. Water (100 mL) is added and stirring is continued for15 minutes. The aqueous phase is extracted thrice with diethyl ether.The combined extracts are washed with saturated aqueous sodiumbicarbonate and saturated aqueous sodium chloride, dried over sodiumsulfate, decanted, and concentrated under reduced pressure to afford 20g of a white paste. The crude material is purified by flashchromatography (ethyl acetate/hexanes) to afford 11 g (56%) of aflocculent white solid. MS (ES⁺): (M+H)=338.1

REFERENCE EXAMPLE 27(4S)-3-[(2S)-2-azido-3-methyl-3-phenylbutanoyl]-4-benzyl-1,3-oxazolidin-2-one

A solution of(4S)-4-benzyl-3-(3-methyl-3-phenylbutanoyl)-1,3-oxazolidin-2-one (1.0 g,3.0 mmol, from Reference Example 26) in anhydrous tetrahydrofuran (16mL) is cooled to −78° C. in a dry-ice/acetone bath while stirring undera nitrogen atmosphere. Potassium hexamethylsilazide (0.5 M solution intoluene, 6.6 mL, 3.3 mmol) is added dropwise to the solution viasyringe. After stirring for 1 hour at −78° C., trisyl azide (1.2 g, 3.8mmol) in tetrahydrofuran (8 mL) is added dropwise. The reaction mixtureis stirred for 2-3 minutes at −78° C. and then quenched by the additionof glacial acetic acid (0.8 mL). Stirring is continued for 1 hour at 40°C. The reaction mixture is diluted with brine and diethyl ether. Theaqueous phase is extracted thrice with diethyl ether. The combinedextracts are washed with saturated aqueous sodium hydrogen carbonate andsaturated aqueous sodium chloride, dried over sodium sulfate, decanted,and concentrated under reduced pressure to afford a white/yellow solid.The crude material is purified by flash chromatography (ethylacetate/hexanes) to afford a clear, straw colored oil (1.1 g, 79%). TOFMS (ES⁺): (M+H)=379.2

REFERENCE EXAMPLE 28 (2S)-2-azido-3-methyl-3-phenylbutanoic acid

2:1 Mixture of S to R

To a 0° C. solution(4S)-3-[(2S)-2-azido-3-methyl-3-phenylbutanoyl]-4-benzyl-1,3-oxazolidin-2-one(0.40 g, 1.1 mmol, from Reference Example 27) in tetrahydrofuran (16 mL)and water (5 mL) is added 30% aqueous hydrogen peroxide (0.26 mL). Afterone to two minutes of stirring, lithium hydroxide monohydrate (92 mg) isadded and the reaction mixture is allowed to stir overnight whilewarming to room temperature.

On the following day, the reaction mixture is cooled to 0° C. andaqueous sodium sulfite solution (1.5 M, 5 mL) is added. After stirringfor an hour at room temperature, the reaction mixture is concentratedunder reduced pressure. The aqueous residue is extracted twice withdichloromethane, then acidified to pH 1 with 10% aqueous hydrochloricacid solution. The acidified aqueous phase is then extracted thrice withethyl acetate. The combined organic extracts are washed with saturatedaqueous sodium chloride, dried over anhydrous sodium sulfate, decanted,and concentrated under reduced pressure to give a clear light blond oil(0.25 mg,>100%). MS (ES⁻): (M−H)=217.9

EXAMPLE 93 Ethyl(E,4S)-4-[((2S)-2-{[(2S)-2-azido-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate and ethyl(E,4S)-4-[((2S)-2-{[(2R)-2-azido-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate

a 2:1 Mixture of S,S,S to R,S,S Diastereomers

To a cooled (0° C., ice bath) solution of a 2:1 mixture of(2S)-2-azido-3-methyl-3-phenylbutanoic acid to(2R)-2-azido-3-methyl-3-phenylbutanoic acid (0.25 g, 1.1 mmol, fromReference Example 28), hydroxybenzotriazole (0.19 g, 1.4 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimine hydrochloride (0.31 g, 1.6mmol) in anhydous dimethylformamide (4 ml) is added N-methylmorpholine(0.19 mL, 1.4 mmol) via syringe under an inert atmosphere. Afterstirring for 15 minutes at 0° C., the cooling bath is removed, and theresulting mixture is stirred for 1 hour. The solution is cooled at 0° C.(ice water bath), and to this mixture is added a solution of ethyl(2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoatehydrochloride (1.1 mmol) in anhydrous dimethylformamide (4 ml). Thecooling bath is removed, and the resulting mixture is stirred for 22hours at room temperature under an inert atmosphere. The mixture isdiluted with water, and the aqueous layer is extracted with ethylacetate (3 times). The combined extracts are washed thrice withsaturated aqueous sodium bicarbonate solution and twice with saturatedaqueous sodium chloride, dried over anhydrous sodium sulfate, filteredand concentrated in vacuo. The residue is chromatographed (silica gel,flash column), to provide a clear, colorless semisolid (0.35 g, 61%). MS(ES⁺): (M+H)=514.0

EXAMPLE 94(E,4S)-4-[((2S)-2-{[(2S)-2-azido-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoicacid and(E,4S)-4-[((2S)-2-{[(2R)-2-azido-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoicacid

A 2:1 mixture of ethyl(E,4S)-4-[((2S)-2-{[(2S)-2-azido-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateand ethyl(E,4S)-4-[((2S)-2-{[(2R)-2-azido-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(0.17 g, 0.35 mmol, from Example 93) is dissolved in methanol (1.5 mL)and tetrahydrofuran (1.5 mL) and cooled to 0° C. (ice water bath). Tothis solution is added water (0.6 mL) and aqueous lithium hydroxidemonohydrate (30 mg, 0.70 mmol). The cooling bath is removed, and theresulting mixture is stirred at 50° C. for 15 hours. Solvents areremoved in vacuo to provide a beige foam, which is then dissolved inwater. The solution is acidified to pH 1 with 2 M aqueous hydrochloricacid solution and then extracted thrice with ethyl acetate. The combinedextracts are washed with saturated sodium chloride solution, dried overanhydrous sodium sulfate, decanted, and concentrated under reducedpressure to afford a hard white foam (0.20 g, 95%). MS (ES⁺):(M+H)=486.0

REFERENCE EXAMPLE 29 2-benzyl-3-methyl-3-phenylbutanoic acid

In accordance with a modified literature procedure (Pfeffer, P. E.;Silbert, L. S.; Chirinko, J. M. J. Org. Chem. 37(3), 1972, 451-458), asolution of 3,3-dimethyl-3-phenyl propionic acid (0.22 g, 1.2 mmol, fromReference Example 22) in tetrahydrofuran (2 mL) is cooled to −40° C.(dry ice/acetone) while stirring under a nitrogen atmosphere. Lithiumdiisopropylamide (Aldrich, 2.0 M solution intetrahydrofuran/ethylbenzene/heptane, 1.4 mL, 2.8 mmol) is addeddropwise via syringe, followed by 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU, 0.16 mL, 1.3 mmol). The reaction mixture is thenallowed to warm to room temperature and to stir for 30 minutes beforebeing re-cooled to 0° C. (ice water bath). Benzyl bromide (0.15 mL, 1.3mmol) is then added neat via syringe and the reaction mixture is allowedto stir overnight while warming to room temperature. The reaction isquenched by the addition of 10% aqueous hydrochloric acid solution andextracted thrice with ethyl acetate. The combined extracts are washedtwice with 10% aqueous hydrochloric acid solution and once withsaturated sodium chloride solution, dried over anhydrous sodium sulfite,and concentrated under reduced pressure. Flash chromatography (silicagel, ether/hexanes) is performed to give a straw colored gum (0.91 g,70% for two combined reactions). MS (ES⁻): (M−H)=266.9

REFERENCE EXAMPLE 30 2-ethyl-3-methyl-3-phenylbutanoic acid

In accordance with a modified literature procedure (Pfeffer, P. E.;Silbert, L. S.; Chirinko, J. M. J. Org. Chem. 37(3), 1972, 451-458), asolution of 3,3-dimethyl-3-phenyl propionic acid (0.47 g, 2.6 mmol, fromReference Example 22) in tetrahydrofuran (4.5 mL) is cooled to −40° C.(dry ice/acetone) while stirring under a nitrogen atmosphere. Lithiumdiisopropylamide (Aldrich, 2.0 M solution intetrahydrofuran/ethylbenzene/heptane, 2.9 mL, 5.8 mmol) is addeddropwise via syringe, followed by 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU, 0.35 mL, 2.9 mmol). The reaction mixture is thenallowed to warm to room temperature and to stir for 30 minutes beforebeing re-cooled to 0° C. (ice water bath). Ethyl iodide (0.31 mL, 3.9mmol) is then added neat via syringe and the reaction mixture is allowedto stir overnight while warming to room temperature. The reaction isquenched by the addition of 10% aqueous hydrochloric acid solution andextracted thrice with ethyl acetate. The combined extracts are washedtwice with 10% aqueous hydrochloric acid solution and once withsaturated sodium chloride solution, dried over anhydrous sodium sulfite,and concentrated under reduced pressure. The crude solid isrecrystallized from hexanes to afford pure material as a white solid(0.16 g, 30%). MS (ES⁻): (M−H)=205.0

REFERENCE EXAMPLE 31 2-allyl-3-methyl-3-phenylbutanoic acid

In accordance with a modified literature procedure (Pfeffer, P. E.;Silbert, L. S.; Chirinko, J. M. J. Org. Chem. 37(3), 1972, 451-458), asolution of 3,3-dimethyl-3-phenyl propionic acid (0.50 g, 2.8 mmol, fromReference Example 22) in tetrahydrofuran (5 mL) is cooled to −40° C.(dry ice/acetone) while stirring under a nitrogen atmosphere. Lithiumdiisopropylamide (Aldrich, 2.0 M solution intetrahydrofuran/ethylbenzene/heptane, 3.1 mL, 6.2 mmol) is addeddropwise via syringe, followed by 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU, 0.37 mL, 4.2 mmol). The reaction mixture is thenallowed to warm to room temperature and to stir for 30 minutes beforebeing re-cooled to 0° C. (ice water bath). Allyl iodide (0.15 mL, 1.3mmol) is then added neat via syringe and the reaction mixture is allowedto stir overnight while warming to room temperature. The reaction isquenched by the addition of 10% aqueous hydrochloric acid solution andextracted thrice with ethyl acetate. The combined extracts are washedtwice with 10% aqueous hydrochloric acid solution and once withsaturated sodium chloride solution, dried over anhydrous sodium sulfite,and concentrated under reduced pressure. Flash chromatography (silicagel, hexane/dichloromethane/ethyl acetate) is performed to give a clearblond oil (0.39 g, 64%). MS (ES⁻): (M−H)=217.0

REFERENCE EXAMPLE 32 3-methyl-2-(methylsulfanyl)-3-phenylbutanoic acid

In accordance with a modified literature procedure (Pfeffer, P. E.;Silbert, L. S.; Chirinko, J. M. J. Org. Chem. 37(3), 1972, 451-458), asolution of 3,3-dimethyl-3-phenyl propionic acid (0.50 g, 2.8 mmol, fromReference Example 22) in tetrahydrofuran (5 mL) is cooled to −40° C.(dry ice/acetone) while stirring under a nitrogen atmosphere. Lithiumdiisopropylamide (Aldrich, 2.0 M solution intetrahydrofuran/ethylbenzene/heptane, 3.1 mL, 6.2 mmol) is addeddropwise via syringe, followed by 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU, 0.37 mL, 3.1 mmol). The reaction mixture is thenallowed to warm to room temperature and to stir for 30 minutes beforebeing re-cooled to 0° C. (ice water bath). Dimethyl disulfide (0.38 mL,4.2 mmol) is then added neat via syringe and the reaction mixture isallowed to stir for two days while warming to room temperature. Thereaction is quenched by the addition of 10% aqueous hydrochloric acidsolution and extracted thrice with ethyl acetate. The combined extractsare washed twice with 10% aqueous hydrochloric acid solution and oncewith saturated sodium chloride solution, dried over anhydrous sodiumsulfite, and concentrated under reduced pressure. Flash chromatography(silica gel, hexanes/dichloromethane/methanol) is performed to give aclear light brown oil which solidified upon standing (0.54 g, 86%). TOFMS (ES⁻): (M−H)=222.9

REFERENCE EXAMPLE 33 3-methyl-2-(methylsulfonyl)-3-phenylbutanoic acid

A solution of 3-methyl-2-(methylsulfanyl)-3-phenylbutanoic acid (0.28 g,1.2 mmol, from Example 32) in dichloromethane (10 mL) is cooled to 0° C.(ice water bath). Peracetic acid (32% aqueous solution, 0.76 mL, 3.6mmol) is added quickly via pipette. The cooling bath is then removed andthe reaction mixture is stirred at room temperature. After three hours,an additional quantity of peracetic acid (32% aqueous solution, 0.76 mL,3.6 mmol) is added. After stirring overnight, the reaction isconcentrated under reduced pressure, while maintaining a bathtemperature of less than 33° C. To the residue, 1.5 M aqueous sodiumsulfite solution (20 mL) is added. The quenched reaction mixture isextracted thrice with ethyl acetate. The combined extracts are washedwith 10% aqueous hydrochloric acid and saturated aqueous sodium chloridesolution, dried over anhydrous sodium sulfate, and concentrated underreduced pressure to give a white powder (0.29 g, 94%). MS (ES⁻):(M−H)=255.0

EXAMPLE 95 Ethyl(E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-(benzyl)-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateSSS Configuration and RSS Configuration

To a solution of 2-benzyl-3-methyl-3-phenylbutanoic acid (0.16 g, 0.60mmol, from Reference Example 29) in anhydrous dichloromethane (2.5 mL)is added triethylethylamine (0.17 mL, 1.2 mmol) under an inertatmosphere. To this solution is added a solution of ethyl(2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoatehydrochloride (0.60 mmol, prepared by using the literature procedures:Ref. Andersen, R, WO 96/33211) in anhydrous dichloromethane (4 mmol),containing triethylamine (0.25 mL, 1.8 mmol). To the mixture of acid andamine is added, bis(2-oxo-3-oxazolidinyl)phosphinic chloride (0.17 g,0.66 mmol). After stirring at room temperature for 15-20 hours, themixture is diluted with water, and the aqueous layer is extracted withethyl acetates (3 times). The combined extracts are washed with 10%aqueous hydrochloric acid, 2 M aqueous sodium carbonate solution, andsaturated aqueous sodium chloride solution, dried over anhydrous sodiumsulfate, filtered and concentrated in vacuo. The residue ischromatographed (silica gel, flash column), to provide a 1:1 mixture oftwo diastereoisomers (SSS and RSS). MS (ES⁺): (M+H)=562.9

EXAMPLE 96(E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-benzyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoicacid SSS Configuration and RSS Configuration

According to General Procedure II, a 1:1 mixture of ethyl(E,4S)-4-[((2S)-2-{[(2S)-2-benzyl-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateand of ethyl(E,4S)-4-[((2S)-2-{[(2R)-2-azido-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(0.17 g, 0.35 mmol, from Example 95) is dissolved in methanol (1.5 mL)and tetrahydrofuran (1.5 mL), and cooled to 0° C. (ice water bath). Tothis solution is added water (0.75 mL) and lithium hydroxide monohydrate(23 mg, 0.54 mmol). The cooling bath is removed, and the resultingmixture is stirred at 60° C. for 16 hours. Solvents are removed in vacuoand the residue is then dissolved in water. The solution is acidified topH 1 with 2 M aqueous hydrochloric acid solution and then extractedthrice with ethyl acetate. The combined extracts are washed withsaturated sodium chloride solution, dried over anhydrous sodium sulfate,decanted, and concentrated under reduced pressure to afford a hard whitefoam, which is crushed to a powder (0.14 g, 100%). MS (ES⁻): (M−H)=533.0

EXAMPLE 97 Ethyl(E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-allyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateSSS Configuration and RSS Configuration

To a solution of 2-allyl-3-methyl-3-phenylbutanoic acid (0.36 g, 1.6mmol, from Reference Example 31) in anhydrous dichloromethane (7 mL) isadded triethylethylamine (0.45 mL, 3.2 mmol) under an inert atmosphere.To this solution is added a solution of ethyl(2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoatehydrochloride (1.6 mmol, prepared by using the literature procedures:Ref. Andersen, R, WO 96/33211) in anhydrous dichloromethane (15 mL),containing triethylamine (0.66 mL, 4.8 mmol). To the mixture of acid andamine is added, bis(2-oxo-3-oxazolidinyl)phosphinic chloride (0.45 g,1.8 mmol). After stirring at room temperature for 16 hours, the mixtureis diluted with water, and the aqueous layer is extracted with ethylacetates (3 times). The combined extracts are washed with 10% aqueoushydrochloric acid, 2 M aqueous sodium carbonate solution, and saturatedaqueous sodium chloride solution, dried over anhydrous sodium sulfate,filtered and concentrated in vacuo. The residue is chromatographed(silica gel, flash column, hexanes/dichloromethane/ethyl acetate), toprovide 0.43 g (52%) of a 1:1 mixture of two diastereoisomers (SSS andRSS). MS (ES⁺): (M+H)=513.0

EXAMPLE 98(E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-allyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoicacid SSS Configuration and RSS Configuration

According to general procedure 11, a 1:1 mixture of ethyl(E,4S)-4-[((2S)-2-{[(2S)-2-allyl-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateand of ethyl(E,4S)-4-[((2S)-2-{[(2R)-2-allyl-3-methyl-3-phenylbutanoyl]amino]-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(0.38 g, 0.74 mmol, from Example 97) is dissolved in methanol (4 mL) andtetrahydrofuran (4 mL), and cooled to 0° C. (ice water bath). To thissolution is added water (2 mL) and lithium hydroxide monohydrate (62 mg,1.5 mmol). The cooling bath is removed, and the resulting mixture isstirred at 60° C. for 15 hours. Solvents are removed in vacuo and theresidue is then dissolved in water. The solution is acidified to pH 1with 2 M aqueous hydrochloric acid solution and then extracted thricewith ethyl acetate. The combined extracts are washed with saturatedsodium chloride solution, dried over anhydrous sodium sulfate, decanted,and concentrated under reduced pressure to afford a hard white foam,which is crushed to a powder (0.32 g, 89%). MS (ES⁺): (M+H)=485.0

EXAMPLE 99 Ethyl(E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-ethyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateSSS Configuration and RSS Configuration

To a solution of 2-ethyl-3-methyl-3-phenylbutanoic acid (0.22 g, 1.1mmol, obtained from Reference Example 30) in anhydrous dichloromethane(5 mL) is added triethylethylamine (0.30 mL, 2.2 mmol) under an inertatmosphere. To this solution is added a solution of ethyl(2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoatehydrochloride (1.1 mmol, prepared by using the literature procedures:Ref. Andersen, R, WO 96/33211) in anhydrous dichloromethane (8 mL),containing triethylamine (0.45 mL, 3.3 mmol). To the mixture of acid andamine is added, bis(2-oxo-3-oxazolidinyl)phosphinic chloride (0.30 g,1.2 mmol). After stirring at room temperature for 16 hours, the mixtureis diluted with water, and the aqueous layer is extracted with ethylacetates (3 times). The combined extracts are washed with 2% aqueoushydrochloric acid, twice with saturated aqueous sodium bicarbonatesolution, and saturated aqueous sodium chloride solution, dried overanhydrous sodium sulfate, filtered and concentrated in vacuo. Theresidue is chromatographed (silica gel, flash column,hexanes/dichloromethane/ethyl acetate), to provide 0.25 g (45%) of a 1:1mixture of two diastereoisomers (SSS and RSS) as a hard white foam,which is crushed to a powder. MS (ES⁺): (M+H)=501.0

EXAMPLE 100(E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-ethyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoicacid SSS Configuration and RSS Configuration

According to General Procedure II, a 1:1 mixture of ethyl(E,4S)-4-[((2S)-2-{[(2S)-2-ethyl-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateand ethyl(E,4S)-4-[((2S)-2-{[(2R)-2-ethyl-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(0.22 g, 0.44 mmol, from Example 99) is dissolved in methanol (2 mL) andtetrahydrofuran (2 mL), and cooled to 0° C. (ice water bath). To thissolution is added water (1 mL) and lithium hydroxide monohydrate (37 mg,0.88 mmol). The cooling bath is removed, and the resulting mixture isstirred at 60° C. for 16 hours. Solvents are removed in vacuo and theresidue is then dissolved in water. The solution is acidified to pH 1with 2 M aqueous hydrochloric acid solution and then extracted thricewith ethyl acetate. The combined extracts are washed with saturatedsodium chloride solution, dried over anhydrous sodium sulfate, decanted,and concentrated under reduced pressure to afford a tan foam (0.21 g,100%). MS (ES⁺): (M+H)=473.0

EXAMPLE 101 Ethyl(E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-methylsulfanyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateSSS Configuration and RSS Configuration

To a solution of 2-methylsulfanyl-3-methyl-3-phenylbutanoic acid (0.25g, 1.1 mmol, obtained from Reference Example 32) in anhydrousdichloromethane (5 mL) is added triethylethylamine (0.30 mL, 2.2 mmol)under an inert atmosphere. To this solution is added a solution of ethyl(2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoatehydrochloride (1.1 mmol, prepared by using the literature procedures:Ref. Andersen, R, WO 96/33211) in anhydrous dichloromethane (8 mL),containing triethylamine (0.45 mL, 3.3 mmol). To the mixture of acid andamine is added, bis(2-oxo-3-oxazolidinyl)phosphinic chloride (0.30 g,1.2 mmol). After stirring at room temperature for 16 hours, the mixtureis diluted with water, and the aqueous layer is extracted with ethylacetates (3 times). The combined extracts are washed with 2% aqueoushydrochloric acid, twice with saturated aqueous sodium bicarbonatesolution, and saturated aqueous sodium chloride solution, dried overanhydrous sodium sulfate, filtered and concentrated in vacuo. Theresidue is chromatographed (silica gel, flash column,hexanes/dichloromethane/ethyl acetate), to provide 0.24 g (42%) of a 1:1mixture of two diastereoisomers (SSS and RSS) as a sticky, tan foam. MS(ES⁺): (M+H)=519.0

EXAMPLE 102(E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-methylsulfanyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoicacid SSS Configuration and RSS Configuration

According to General Procedure II, a 1:1 mixture of ethyl(E,4S)-4-[((2S)-2-{[(2S)-2-methylsulfanyl-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateand ethyl(E,4S)-4-[((2S)-2-{[(2R)-2-methylsulfanyl-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(0.20 g, 0.39 mmol, from Example 101) is dissolved in methanol (2 mL)and tetrahydrofuran (2 mL), and cooled to 0° C. (ice water bath). Tothis solution is added water (1 mL) and lithium hydroxide monohydrate(33 mg, 0.78 mmol). The cooling bath is removed, and the resultingmixture is stirred at 60° C. for 15 hours. Solvents are removed in vacuoand the residue is then dissolved in water. The solution is acidified topH 1 with 2 M aqueous hydrochloric acid solution and then extractedthrice with ethyl acetate. The combined extracts are washed withsaturated sodium chloride solution, dried over anhydrous sodium sulfate,decanted, and concentrated under reduced pressure to afford a light tanpowder (0.20 g,>100%). MS (ES⁺): (M+H)=490.9

REFERENCE EXAMPLE 34 Ethyl(E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-methylsulfonyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateSSS Configuration and RSS Configuration

To a solution of 2-methylsulfonyl-3-methyl-3-phenylbutanoic acid (0.27g, 1.1 mmol, from Reference Example 33) in anhydrous dichloromethane (5mL) is added triethylethylamine (0.30 mL, 2.2 mmol) under an inertatmosphere. To this solution is added a solution of ethyl(2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoatehydrochloride (1.1 mmol, prepared by using the literature procedures:Ref. Andersen, R, WO 96/33211) in anhydrous dichloromethane (8 mL),containing triethylamine (0.45 mL, 3.3 mmol). To the mixture of acid andamine is added, bis(2-oxo-3-oxazolidinyl)phosphinic chloride (0.30 g,1.2 mmol). After stirring at room temperature for 16 hours, the mixtureis diluted with water, and the aqueous layer is extracted with ethylacetates (3 times). The combined extracts are washed with 2% aqueoushydrochloric acid, twice with saturated aqueous sodium bicarbonatesolution, and saturated aqueous sodium chloride solution, dried overanhydrous sodium sulfate, filtered and concentrated in vacuo. Theresidue is purified by reverse-phase HPLC (water/acetonitrile) toprovide 0.20 g (33%) of a 1:1 mixture of two diastereoisomers (SSS andRSS) as a white powder. MS (ES⁺): (M+H)=551.3

EXAMPLE 103(E,4S)-4-[((2S)-3,3-dimethyl-2-{[3-methyl-2-methylsulfonyl-3-phenylbutanoyl]amino}butanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoicacid SSS Configuration and RSS Configuration

According to General Procedure II, a 1:1 mixture of ethyl(E,4S)-4-[((2S)-2-{[(2S)-2-methylsulfonyl-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateand ethyl(E,4S)-4-[((2S)-2-{[(2R)-2-methylsulfonyl-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(0.17 g, 0.31 mmol, from Reference Example 34) is dissolved in methanol(1.5 mL) and tetrahydrofuran (1.5 mL), and cooled to 0° C. (ice waterbath). To this solution is added water (0.75 mL) and lithium hydroxidemonohydrate (26 mg, 0.62 mmol). The cooling bath is removed, and theresulting mixture is stirred at 55-60° C. for 16 hours. Solvents areremoved in vacuo and the residue is then dissolved in water. Thesolution is acidified to pH 1 with 2 M aqueous hydrochloric acidsolution and then extracted thrice with ethyl acetate. The combinedextracts are washed with saturated sodium chloride solution, dried overanhydrous sodium sulfate, decanted, and concentrated under reducedpressure to afford a hard white foam, which is crushed to a powder (0.17g, >100%). TOF MS (ES⁻): (M−H)=521.0

REFERENCE EXAMPLE 35 3-phenyl-3-methyl-2-oxobutanoic acid

To a 500 mL 3-neck round-bottom flask equipped with a reflux condenserand magnetic stirrer containing 3-phenyl-2-oxopropanoic acid (25 g, 150mmol) as a solution in tetrahydrofuran (130 mL) and water (50 mL) isadded 5 M sodium hydroxide solution (80 mL) and iodomethane (25 mL, 400mmol). While stirring under a nitrogen atmosphere, the reaction mixtureis heated in a 68-71° C. oil bath for 5½ hours. After this interval hadpassed, additional volumes of 5 N NaOH (50 mL) and iodomethane (10 mL)are added. The reaction mixture is allowed to stir for 2 days at roomtemperature, then volatiles are evaporated under reduced pressure. Theaqueous residue is extracted thrice with ethyl acetates and thenacidified to pH 1 by the addition of concentrated hydrochloric acid. Theacidified phase is extracted thrice with ethyl acetates. The combinedextracts are washed twice with saturated aqueous sodium chloridesolution, dried over anhydrous sodium sulfate, decanted, andconcentrated under reduced pressure to afford a dark burgundy liquid (31g,>100%). MS (ES⁻): (M−H)=190.8

REFERENCE EXAMPLE 36 Methyl 3-phenyl-3-methyl-2-oxobutanoate

To a 0° C. (ice water bath) solution of 3-phenyl-3-methyl-2-oxobutanoate(9.0 g, 47 mmol, from Reference Example 35) in ether (40 mL) andmethanol (60 mL). Trimethylsilyl diazomethane (2.0 M solution inhexanes, 35 mL, 70 mmol, Aldrich) is added dropwise until the evolutionof gas ceased and the yellow color of the reagent persisted in solution.The solvents are evaporated under reduced pressure and the residue istaken up in ether/hexanes (1:1) and washed with 2% aqueous hydrochloricacid, saturated aqueous sodium bicarbonate solution, and saturatedaqueous sodium chloride solution. The combined extracts are then driedover anhydrous sodium sulfate, and concentrated under reduced pressureto give a straw-colored liquid (8.5 g, 88%). MS (ES⁺): (M+H)=206.9

REFERENCE EXAMPLE 37 Methyl 2-hydroxy-3-methyl-3-phenylbutanoate

Following a procedure found in the literature (Ferguson, C. G.; Money,T.; Pontillo, J.; Whitelaw, P. D. M.; Wong, M. K. C. Tetrahedron.52(47), 1996, 14661-14627) a solution of methyl3-phenyl-3-methyl-2-oxobutanoate (2.5 g, 12 mmol, from Reference Example36) in methanol (144 mL) is cooled to 0° C. (ice water bath). Sodiumborohydride (0.21 g, 5.5 mmol) is added in three portions over 3minutes. The reaction mixture is stirred at 0° C. for 10 minutes andthen at room temperature for 30 minutes. The reaction mixture isquenched by the addition of 10% aqueous hydrochloric acid solution untilpH=6 and then extracted thrice with diethyl ether. The combined extractsare washed with water and saturated aqueous sodium chloride solution,dried over anhydrous sodium sulfite, and concentrated under reducedpressure to give a light blond liquid, which is combined with the crudematerial from another reaction. The combined crude material is subjectedto flash chromatography to (silica gel, hexanes/ethyl acetate) andprovided 2.4 g (80% combined yield) of a clear, colorless oil. TOF MS(ES⁺): (M+H)=209.1

REFERENCE EXAMPLE 38 Methyl 2-methoxy-3-methyl-3-phenylbutanoate

To a solution of methyl 2-hydroxy-3-methyl-3-phenylbutanoate (0.50 g,2.4 mmol, from Reference Example 37) in diethyl ether (11 mL) is addedsilver (I) oxide (4.3 g) and iodomethane (8.7 mL). The reaction mixtureis stirred in a 40° C. oil bath under nitrogen atmosphere for 26 hours.The reaction mixture is combined with that from a previous run conductedunder the same conditions and filtered through a plug of diatomaceousearth. The filtrate is concentrated under reduced pressure and thenpurified by flash chromatograhy (silica gel, hexanes/ethyl acetate) tofurnish 0.52 g of a clear, colorless liquid (68% combined yield). MS(ES⁺): (M+H)=222.9

REFERENCE EXAMPLE 39 2-methoxy-3-methyl-3-phenylbutanoic acid

According to General Procedure II, methyl2-methoxy-3-methyl-3-phenylbutanoic acid (0.49 g, 2.2 mmol, fromReference Example 38) is dissolved in methanol (10 mL) andtetrahydrofuran (10 mL). To this solution is added water (5 mL) andlithium hydroxide monohydrate (0.18 g, 4.4 mmol), and the resultingmixture is stirred at 60° C. for 16 hours. Solvents are removed in vacuoand the residue is then dissolved in water. The solution is acidified topH 1 with 10% aqueous hydrochloric acid solution and then extractedthrice with ethyl acetate. The combined extracts are washed withsaturated sodium chloride solution, dried over anhydrous sodium sulfate,decanted, and concentrated under reduced pressure to afford a lightorange oil, which solidified to a straw colored crystalline solid (0.46g, 100%). MS (ES⁻): (M−H)=207.0

EXAMPLE 104 Ethyl(E,4S)-4-[{(2S)-2-[(2-methoxy-3-methyl-3-phenylbutanoyl)amino]-3,3-dimethylbutanoyl}(methyl)amino]-2,5-dimethyl-2-hexenoateSSS Configuration and RSS Configuration

To a solution of 2-methoxy-3-methyl-3-phenylbutanoic acid (0.43 g, 2.1mmol, from Reference Example 39), hydroxybenzotriazole (0.33 g, 2.5mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimine hydrochloride(0.55 g, 2.9 mmol) in anhydrous dimethylformamide (7 ml) is addedN-methylmorpholine (0.34 mL, 3.1 mmol) via syringe under an inertatmosphere. After stirring for 1 hour at room temperature, a solution ofethyl (2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoatehydrochloride (2.1 mmol) is added. The resulting mixture is stirred for70 hours at room temperature under an inert atmosphere. The mixture isdiluted with water, and the aqueous layer is extracted with diethylether (3 times). The combined extracts are washed with 2% aqueoushydrochloric acid (×2), saturated aqueous sodium bicarbonate solution(×2), water, and saturated aqueous sodium chloride. The extracts aredried over anhydrous sodium sulfate, filtered and concentrated in vacuoto give a 1:1 SSS to RSS mixture of ethyl(E,4S)-4-[{(2S)-2-[(2-methoxy-3-methyl-3-phenylbutanoyl)amino]-3,3-dimethylbutanoyl}(methyl)amino]-2,5-dimethyl-2-hexenoateas a pale straw colored gum (1.0 g, 91%). MS (ES⁺): (M+H)=503.0

EXAMPLE 105(E,4S)-4-[{N-[(2S)-2-methoxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoicacid

According to General Procedure II, a 1:1 mixture of ethyl(E,4S)-4-[((2S)-2-{[(2S)-2-methoxy-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateand ethyl(E,4S)-4-[((2S)-2-{[(2R)-2-methoxy-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(1.0 g, 2.0 mmol, from Reference Example 104) is dissolved in methanol(10 mL) and tetrahydrofuran (10 mL). To this solution is added water (5mL) and lithium hydroxide monohydrate (0.17 g, 4.0 mmol), and theresulting mixture is stirred at 60° C. for 16 hours. Solvents areremoved in vacuo and the residue is then dissolved in water. Thesolution is acidified to pH 1 with 2 M aqueous hydrochloric acidsolution and then extracted thrice with ethyl acetate. The combinedextracts are washed with saturated sodium chloride solution, dried overanhydrous sodium sulfate, decanted, and concentrated under reducedpressure to afford a white foam. A sample of this crude material (240mg) is subjected to HPLC purification (Prodigy OS3 column,acetonitrile/water/TFA) to give(E,4S)-4-[{N-[(2S)-2-methoxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoicacid as a hard white foam (50 mg). MS (ES⁺): (M+H)=475.0

EXAMPLE 106(E,4S)-4-[{N-[(2R)-2-methoxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoicacid

According to General Procedure II, a 1:1 mixture of ethyl(E,4S)-4-[((2S)-2-{[(2S)-2-methoxy-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateand ethyl(E,4S)-4-[((2S)-2-{[(2R)-2-methoxy-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(1.0 g, 2.0 mmol, from Reference Example 104) is dissolved in methanol(10 mL) and tetrahydrofuran (10 mL). To this solution is added water (5mL) and lithium hydroxide monohydrate (0.17 g, 4.0 mmol), and theresulting mixture is stirred at 60° C. for 16 hours. Solvents areremoved in vacuo and the residue is then dissolved in water. Thesolution is acidified to pH 1 with 2 M aqueous hydrochloric acidsolution and then extracted thrice with ethyl acetate. The combinedextracts are washed with saturated sodium chloride solution, dried overanhydrous sodium sulfate, decanted, and concentrated under reducedpressure to afford a white foam. A sample of this crude material (240mg) is subjected to HPLC purification (Prodigy OS3 column,acetonitrile/water/TFA) to give(E,4S)-4-[(N-[(2R)-2-methoxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoicacid as a clear colorless oil (80 mg). MS (ES⁺): (M+H)=475.0

REFERENCE EXAMPLE 40 2-hydroxy-3-methyl-3-phenylbutanoic acid

According to general procedure, methyl2-hydroxy-3-methyl-3-phenylbutanoic acid (0.27 g, 1.3 mmol, fromReference Example 37) is dissolved in methanol (6 mL) andtetrahydrofuran (6 mL). To this solution is added water (3 mL) andlithium hydroxide monohydrate (0.12 g, 2.9 mmol), and the resultingmixture is stirred at 60-65° C. for 16 hours. Solvents are removed invacuo and the residue is then dissolved in water. The solution isacidified to pH 1 with 2 M aqueous hydrochloric acid solution and thenextracted thrice with ethyl acetate. The combined extracts are washedwith saturated sodium chloride solution, dried over anhydrous sodiumsulfate, decanted, and concentrated under reduced pressure to afford alight pink oil, which solidified under vacuum (0.25 g, 100%). TOF MS(ES⁻): (M−H)=193.0

REFERENCE EXAMPLE 41 Ethyl(E,4S)-4-[{(2S)-2-[(2-hydroxy-3-methyl-3-phenylbutanoyl)amino]-3,3-dimethylbutanoyl}(methyl)amino]-2,5-dimethyl-2-hexenoateSSS Configuration and RSS Configuration

To a solution of 2-hydroxy-3-methyl-3-phenylbutanoic acid (0.23 g, 1.2mmol, from Reference Example 40), hydroxybenzotriazole (0.19 g, 1.4mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimine hydrochloride(0.33 g, 1.7 mmol) in anhydrous dimethylformamide (4 ml) is addedN-methylmorpholine (0.20 mL, 1.8 mmol) via syringe under an inertatmosphere. After stirring for 1 hour at room temperature, a solution ofethyl (2E,4S)-2,5-dimethyl-4-[methyl(3-methyl-L-valyl)amino]hex-2-enoatehydrochloride (1.2 mmol) in anhydrous dimethylformamide (4 ml) is added.The resulting mixture is stirred for 15 hours at room temperature underan inert atmosphere. The mixture is diluted with water, and the aqueouslayer is extracted with diethyl ether (3 times). The combined extractsare washed thrice with 2% aqueous hydrochloric acid, saturated aqueoussodium bicarbonate solution, and saturated aqueous sodium chloride. Theextracts are dried over anhydrous sodium sulfate, filtered andconcentrated in vacuo to give a white powder (0.39 g, 66%). MS (ES⁺):(M+H)=489.6

EXAMPLE 107 AND EXAMPLE 108(E,4S)-4-[{(2S)-2-[(2-hydroxy-3-methyl-3-phenylbutanoyl)amino]-3,3-dimethylbutanoyl}(methyl)amino]-2,5-dimethyl-2-hexenoicacid SSS Configuration and RSS Configuration EXAMPLE 107(E,4S)-4-[{N-[(2S)-2-hydroxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoicacid

A 1:1 mixture of ethyl(E,4S)-4-[((2S)-2-{[(2S)-2-hydroxy-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateand ethyl(E,4S)-4-[((2S)-2-{[(2R)-2-hydroxy-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(0.34 g, 0.70 mmol, from Reference Example 41) is dissolved in methanol(3.2 mL) and tetrahydrofuran (3.2 mL). To this solution is added water(1.6 mL) and lithium hydroxide monohydrate (64 mg, 1.5 mmol), and theresulting mixture is stirred at 65° C. for 16 hours. Solvents areremoved in vacuo and the residue is then dissolved in water. Thesolution is acidified to pH 1 with 2 M aqueous hydrochloric acidsolution and then extracted thrice with ethyl acetate. The combinedextracts are washed with saturated sodium chloride solution, dried overanhydrous sodium sulfate, decanted, and concentrated under reducedpressure to afford a white foam. A sample of this crude material (200mg) is subjected to HPLC purification (Prodigy OS3 column,acetonitrile/water/TFA) to give(E,4S)-4-[{N-[(2S)-2-hydroxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoicacid as a hard white foam, which is crushed into a powder (50 mg). MS(ES⁺): (M+H)=461.0

EXAMPLE 108(E,4S)-4-[{N-[(2R)-2-hydroxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoicacid

A 1:1 mixture of ethyl(E,4S)-4-[((2S)-2-{[(2S)-2-hydroxy-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoateand ethyl(E,4S)-4-[((2S)-2-{[(2R)-2-hydroxy-3-methyl-3-phenylbutanoyl]amino}-3,3-dimethylbutanoyl)(methyl)amino]-2,5-dimethyl-2-hexenoate(0.34 g, 0.70 mmol, from Reference Example 41) is dissolved in methanol(3.2 mL) and tetrahydrofuran (3.2 mL). To this solution is added water(1.6 mL) and lithium hydroxide monohydrate (64 mg, 1.5 mmol), and theresulting mixture is stirred at 65° C. for 16 hours. Solvents areremoved in vacuo and the residue is then dissolved in water. Thesolution is acidified to pH 1 with 2 M aqueous hydrochloric acidsolution and then extracted thrice with ethyl acetate. The combinedextracts are washed with saturated sodium chloride solution, dried overanhydrous sodium sulfate, decanted, and concentrated under reducedpressure to afford a white foam. A sample of this crude material (200mg) is subjected to HPLC purification (Prodigy OS3 column,acetonitrile/water/TFA) to give(E,4S)-4-[{N-[(2R)-2-hydroxy-3-methyl-3-phenylbutanoyl]-3-methyl-L-valyl}(methyl)amino]-2,5-dimethyl-2-hexenoicacid as a white foam, which is crushed to a powder (50 mg). MS (ES⁺):(M+H)=461.0

EXAMPLE 109(2S)-N-[(1S,2E)-4-hydrazino-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N,3,3-trimethyl-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamide

To 150 mg of4-[[(2S)-3,3-dimethyl-2-[[(2S)-3-methyl-2-(methylamino)-1-oxo-3-phenylbutyl]amino)]-1-oxobutyl]methylamino]-2,5-dimethyl-(E,4S)-2-hexenoic acid in 25 mL of acetonitrile at 0° C. is added 95 mg ofN-hydroxysuccinimide and 150 mg of1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride. Thisreaction is allowed to stir overnight at ambient temperature beforeadding 200 uL of hydrazine hydrate in 5 mL of methanol. After anadditional 3 hours at ambient temperature the reaction is poured into amixture of brine and saturated aqueous sodium bicarbonate, and thisaqueous mixture is extracted three times with chloroform. The combinedchloroform is dried with magnesium sulfate and purified by flashchromatography on silica gel with a gradient of 0-4% methanol inchloroform. The concentrated, pooled fractions containing product areprecipitated from ether into hexanes and then filtered and dried to give91 mg of(2S)-N-[(1S,2E)-4-hydrazino-1-isopropyl-3-methyl-4-oxo-2-butenyl]-N,3,3-trimethyl-2-{[(2S)-3-methyl-2-(methylamino)-3-phenylbutanoyl]amino}butanamideas an off-white powder and 37 mg of the same product from theconcentrated mother liquor as a yellowish solid foam (83% total yield).HRMS (ESI): (M+H)=488.35952 (−0.34 mmu).

1. A compound represented by Formula I:

wherein: A is selected from the group consisting of an alkyl moiety of 1to 10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and acyclic hydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atomsmay optionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygenatoms, and 0 to 4 sulfur atoms, and the carbon atoms are optionallysubstituted with: ═O, ═S, —OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀,—NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, —Br, —Cl, —F, —CN,—CO₂H, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂,—SO₃H, —SOR₁₀, —SO₂R₁₀, wherein R₁₀ is an alkyl moiety of 1 to 10 carbonatoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and a cyclichydrocarbon moiety of 3 to 10 carbon atoms, aryl-R— and heteroaryl-R; orA is OR, S(O)R, S(O)₂R, SO₂NR₂, NR₁R₂ or N₃; R is selected from thegroup consisting of H, an alkyl moiety of 1 to 18 carbon atoms, alkenylmoiety of 2 to 18 carbon atoms, aryl and a cyclic hydrocarbon moiety of3 to 18 carbon atoms, wherein carbon atoms may optionally be replacedwith 0 to 4 nitrogen atoms, 0 to 4 oxygen atoms, and 0 to 4 sulfuratoms, and the carbon atoms are optionally substituted with: ═O, ═S,—OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂,—NHCOR₁₀, —NR₁₀COR₁₀, —I, —Br, —Cl, —F, —CN, —CO₂H, —CO₂R₁₀, —CHO,—COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H,—SOR₁₀, or —SO₂R₁₀; B′ is O or H₂; E is the moiety

or aryl, a 5 to 14-membered monocydic, bicyclic or tricyclic saturatedor unsaturated hydrocarbon ring moiety wherein carbon atoms mayoptionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygen atoms,and 0 to 4 sulfur atoms wherein the carbon atoms may optionally besubstituted with: R, ═O, ═S, —OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀,—NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, —Br, —Cl, —F, —CN,—CO₂H, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂,—SO₃H, —SOR₁₀, or —SOR₁₀; R₁ is selected fmm the group consisting of H,an alkyl moiety of 1 to 10 carbon atoms, alkenyl moiety of 2 to 10carbon atoms, aryl and a cyclic hydrocarbon moiety of 3 to 10 carbonatoms, wherein carbon atoms may optionally be replaced with 0 to 4nitrogen atoms, 0 to 4 oxygen atoms, and 0 to 4 sulfur atoms, and thecarbon atoms may optionally be substituted with: ═O, ═S, —OH, —OR₁₀,—O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀,—NR₁₀COR₁₀, —I, —Br, —Cl, —F, —CN, —CO₂H, —CHO, —COR₁₀, —CONH₂,—CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, or —SOR₁₀; R₂is selected from the group consisting of H, an alkyl moiety of 1 to 10carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and a cyclichydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atoms mayoptionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygen atoms,and 0 to 4 sulfur atoms, and the carbon atoms may optionally besubstituted with: ═O, ═S, —OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀,—NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, —Br, —Cl, —F, —ON,—CO₂H, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂,—SO₃H, —SOR₁₀, or —SOR₁₀; R₁ and R₂ together may optionally form a ringof 3 to 7 carbon atoms wherein carbon atoms may optionally be replacedwith 0 to 2 nitrogen atoms, 0 to 2 oxygen atoms and 0 to 2 sulfur atoms;R₃ is selected from the group consisting of H, an alkyl moiety of 1 to10 carbon atoms, alkenyl moiety, of 2 to 10 carbon atoms, aryl and acyclic hydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atomsmay optionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygenatoms, and 0 to 4 sulfur atoms, and the carbon atoms may optionally besubstituted with: ═O, ═S, —OH, —OR₁₀, —O₂CR₁₀, —SH, —SOCR₁₀, —NH₂,—NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, —Br, —Cl, —F, —ON, —CO₂H,—CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀,—NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀; R₄ is selected from the groupconsisting of H, an alkyl moiety of 1 to 10 carbon atoms, alkenyl moietyof 2 to 10 carbon atoms, aryl and a cyclic hydrocarbon moiety of 3 to 10carbon atoms, wherein carbon atoms may optionally be replaced with 0 to4 nitrogen atoms, 0 to 4 oxygen atoms, and 0 to 4 sulfur atoms, and thecarbon atoms may optionally be substituted with: ═O, ═S, —OH, —OR₁₀,—O₂CR₁₀, —SH, —SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I,—Br, —CI, —F, —CN, —CO₂H, —CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀,—CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, or —S₂R₁₀; R₃ and R₄together may optionally form a ring of 3 to 7 carbon atoms whereincarbon atoms may optionally be replaced with 0 to 2 nitrogen atoms, 0 to2 oxygen atoms and 0 to 2 sulfur atoms; R₅ is selected from the groupconsisting of H, OH, NHR, SH, aryl, heteroaryl, an alkyl moiety of 1 to10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms and a cyclichydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atoms mayoptionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygen atoms,and 0 to 4 sulfur atoms, and the carbon atoms may optionally besubstituted with: ═O, ═S, —OH, —OR₁₀, —O₂CR₁₀, —SH, —SOCR₁₀, —NH₂,—NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, Br, —Cl, —F, —CN, —CO₂H,—CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀,—NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀; R₅ and A may optionally form a ring of5 to 7 carbon atoms wherein carbon atoms may optionally be replaced with0 to 2 nitrogen atoms, 0 to 2 oxygen atoms, and 0 to 2 sulfur atoms; R₆is selected from the group consisting of H, an alkyl moiety of 1 to 10carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and a cyclichydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atoms mayoptionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygen atoms,and 0 to 4 sulfur atoms, and the carbon atoms may optionally besubstituted with: ═O, ═S, —OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀,—NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, —Br, —Cl, —F, —CN,—CO₂H, —CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH,—COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, or —SOR₁₀; R₇ is selected from the groupconsisting of H, an alkyl moiety of 1 to 10 carbon atoms, alkenyl moietyof 2 to 10 carbon atoms, aryl and a cyclic hydrocarbon moiety of 3 to 10carbon atoms, wherein carbon atoms may optionally be replaced with 0 to4 nitrogen atoms, 0 to 4 oxygen atoms, and 0 to 4 sulfur atoms, and thecarbon atoms may optionally be substituted with: ═O, ═S, —OH, —OR₁₀,—O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀, —NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀,—NR₁₀COR₁₀, —I, Br, —Cl, —F —CN, —CO₂H, —CO₂R₁₀, —CHO, —COR₁₀, —CONH₂,—CONHR₁₀, —CON(R₁₀)₂, —COSH, —COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀;R₈ is selected from the group consisting of H, an alkyl moiety of 1 to10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryl and acyclic hydrocarbon moiety of 3 to 10 carbon atoms, wherein carbon atomsmay optionally be replaced with 0 to 4 nitrogen atoms, 0 to 4 oxygenatoms, and 0 to 4 sulfur atoms, and the carbon atoms may optionally besubstituted with: ═O, ═S, —OH, —OR₁₀, —O₂CR₁₀, —SH, —SR₁₀, —SOCR₁₀,—NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, —Br, —CI, —F —CN,—CO₂H, —CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH,—COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀;

R₉ is selected from D is O or OH; Y is an alkyl moiety of 1 to 10 carbonatoms optionally substituted with R, AryIR-, or X, or an alkenyl moietyof 2 to 10 carbon atoms optionally substituted with R, ArylR-, or X; Zis defined as a moiety selected from the group consisting of:pyrrolidinyl, pyrrolyl and a moiety of formula

the dotted line is an optional bond; m is 1; R₁₈ is H or OH; R₁₉ isselected from a bond, an alkyl moiety of 1 to 10 carbon atoms optionallysubstituted with an alkyl moiety of 1 to 10 carbon atoms, and alkoxy of1 to 10 carbon atoms; R₂₀ is selected from OR₁₄, NH—R₂₁, a moiety of theformula

and a moiety of the formula

R₂₁ is an alkyl moiety of 1 to 10 carbon atoms optionally substitutedwith aryl and heteroaryl; R₂₂ is an alkyl moiety of 1 to 10 carbon atomsoptionally substituted with aryl; X is defined as a moiety selected fromthe group consisting of: —OH, —OR, ═O, ═S, —O₂CR, —SH, —SR, —SOCR, —NH₂,—NHR, —N(R)₂, —NHCOR, —NRCOR, —I, —Br, —Cl, —F, —CN, —CO₂H, —CO₂R, —CHO,—COR, —CONH₂, —CONHR, —CON(R)₂, —COSH, —COSR, —NO₂, —SO₃H, —SOR, and—SO₂R; Aryl is defined as an aromatic hydrocarbon moiety having 6, 10 or14 carbon atoms, optionally substituted with R or Z; Heteroaryl means a5- or 6-membered heterocyclic ring, which may be fused to another 5- or6-membered heterocyclic ring, especially heteroaromatic rings whichcontain 1 to 3 heteroatoms independently selected from 0, N and Soptionally substituted with R or X or fused to a cyclic hydrocarbonmoiety of 3 to 10 carbon atoms; or pharmaceutically acceptable saltsthereof.
 2. A compound according to claim 1 wherein: (a) R₁ is H,methyl, ethyl, propyl, or n-butyl and R₂ is methyl, ethyl, propyl, orn-butyl; or (b) where R₁ and R₂ taken together with the nitrogen atom towhich they are attached form a three to six membered ring.
 3. A compoundaccording to claim 2 wherein R₁ is H and R₂ is CH₃ .
 4. A compoundaccording to claim 1 wherein no more than one of R₃ and R₄ is H.
 5. Acompound according to claim 1 wherein R₃ and R₄ are independently:methyl, ethyl, n-propyl or n-butyl.
 6. A compound according to claim 1wherein R₃ and R₄ are each methyl.
 7. A compound according to claim 1wherein R₃ and R₄ are joined together to form a β-cyclopropyl,β-cyclobutyl, β-cyclopentyl or β-cyclohexyl ring.
 8. A compoundaccording to claim 1 wherein R₅ is aryl.
 9. A compound according toclaim 1 wherein R₅ is phenyl, or naphthyl.
 10. A compound according toclaim 1 wherein R₅ is phenyl.
 11. A compound according to claim 1wherein R₅ is cyclohexyl.
 12. A compound according to claim 1 wherein Eis phenyl, adamantly, or 2,3,4-tetrahydro-1 -naphthalenyl.
 13. Acompound according to claim 1 wherein R₈ is H and R₈ is methyl.
 14. Acompound according to claim 1 wherein R₇ is a three to six carbon,branched alkyl group.
 15. A compound according to claim 14 wherein R₇ is—C(CH₃)₃.
 16. A compound according to claim 1 wherein R₉ is—C(R₁₅)—C═C(R₁₆)C(O)-Z wherein R₁₅ is methyl, ethyl, n-propyl,isopropyl, tert-butyl, iso-butyl, or sec-butyl and R₁₆ is H, methyl,ethyl, propyl, iso-propyl, n-butyl, iso-butyl or sec-butyl.
 17. Acompound according to claim 1 wherein R₉ is:


18. A compound according to claim 16 wherein R₉ is


19. A compound according to claim 17 wherein R₉ is


20. A compound according to claim 1 wherein: A is NR₁R₂; R₁ is methyl;R₂ is H; R₃ and R₄ are methyl; R₅ is selected from the group consistingof H, an alkyl moiety of 1 to 10 carbon atoms, aryl and a cyclichydrocarbon moiety of 3 to 10 carbon atoms, said carbon atoms beingoptionally substituted with: ═O, ═S, —OH, —OR₁₀, —O₂CR₁₀, —SH, —SOCR₁₀,—NH₂, —NR₁₀H, —N(R₁₀)₂, —NHCOR₁₀, —NR₁₀COR₁₀, —I, —Br, —CI, —F, —CN,—CO₂H, —CO₂R₁₀, —CHO, —COR₁₀, —CONH₂, —CONHR₁₀, —CON(R₁₀)₂, —COSH,—COSR₁₀, —NO₂, —SO₃H, —SOR₁₀, or —SO₂R₁₀, wherein R₁₀ is an alkyl moietyof 1 to 10 carbon atoms, alkenyl moiety of 2 to 10 carbon atoms, aryland a cyclic hydrocarbon moiety of 3 to 10 carbon atoms, aryl-R— andheteroaryl-R; R₇ is t-butyl; R₉ is —C(R₁₅)—C═C(R₁₆)C(O)-Z wherein R₁₅ ismethyl, ethyl, n-propyl, isopropyl, tert-butyl, iso-butyl, or sec-butyland R₁₆ is H, methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl orsec-butyl; or pharmaceutically acceptable salts thereof.
 21. A compoundaccording to claim 20 wherein: R₉ is:


22. A compound according to claim 21 wherein: R₉ is


23. A compound according to claim 22 wherein: R₉ is


24. A compound according to claim 1 wherein the absolute configurationsof moieties a, b and c of Formula (I)

are selected from: a b c S S S R S S and S S R.


25. The compound according to claim 1, N,β,β,3,4-Pentamethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 26. The compoundaccording to claim 1, N, β,β,3,4-Pentamethyl-L-phenylalanyl-N¹-{( 1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 27. The compoundaccording to claim 1, N,β,β,3,5Pentamethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 28. The compoundaccording to claim 1, N,ββ,3,5-Pentamethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 29. The compoundaccording to claim 1, N,ββ,3-Tetramethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 30. The compoundaccording to claim 1, N,ββ,3-Tetramethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 31. The compoundaccording to claim 1, 3-Chloro-N, ββ-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 32. The compoundaccording to claim 1, 3-Chloro-N,ββ-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 33. The compoundaccording to claim 1, N,ββ-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-1-isopropyl-4-[(2S)-2-((1R,2R)-1-methoxy-2-methyl-3-oxo-3{[(1S)-2phenyl-1-(1,3-thiazolyl-2-yl)ethyl]amino}propyl)pyrrolidin-1-yl]-3-methyl-4-oxobut-2-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 34. The compoundaccording to claim 1, N,ββ-trimethyl-L-phenylalanyl-N¹-((1S,2E)-4-{(2S)-2-[(1R,2R)-1,3-dimethoxy-2-methyl-3-oxopropyl]pyrrolidin-1-yl}-1-isopropyl-3-methyl-4-oxobut-2-enyl)-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 35. The compoundaccording to claim 1,N-methyl-3-phenyl-L-valyl-N¹-[(1S,2E)-1-isopropyl-4-((2S)-2-{(1R,2R)-1-methoxy-2-methyl-3-oxo-[(2-phenethyl)amino]propyl}pyrrolidin-1-yl)-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 36. The compoundaccording to claim 1, N,ββ-trimethyl-L-phenylalanyl—N¹-{(1S,2E)-4-[(2S,4R)-2-(methoxycarbonyl)-4-hydroxypyrrolidin-1-yl]-1-isopropyl-3methyl-4oxobut-2-enyl}-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 37. The compoundaccording to claim 1, N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2R)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 38. The compoundaccording to claim 1, N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S,4R)-2-carboxy4-hydroxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 39. The compoundaccording to claim 1, N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2R)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 40. The compoundaccording to claim 1,3-cyclohexyl-N-methyl-L-valyl-N¹-{(1S,2E)-1-isopropyl-4-[(2S)-2-((1R,2R)-1-methoxy-2-methyl-3-oxo-3-{[(1S)-2phenyl-1-(1,3-thiazol-2-yl)ethyl]aminol}propyl)pyrrolidin-1-yl]-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamide or a pharmaceutically acceptable salt thereof.41. The compound according to claim 1,3-cyclohexyl-N-methyl-L-valyl-N1-{(1S,2E)-4-[(2S)-2-(methoxycarbonyl)pyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobutenyl}—N1,3-dimethyl-L-valinamide or a pharmaceutically acceptable saltthereof.
 42. The compound according to claim 1,3-cyclohexyl—N-methyl-L-valyl-N¹-{(1S,2E)-4-[(2S)-2-carboxypyrrolidin-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 43. The compoundaccording to claim 1, N,β,β-trimethyl-L-phenylalanyl-N¹-{(1S,2E)-4-[(2S)-2-carboxy-2,5-dihydro-1H-pyrrol-1-yl]-1-isopropyl-3-methyl-4-oxobut-2-enyl}N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 44. The compoundaccording to claim 1,3-cyclohexyl-N-methyl-L-valyl-N¹-[(1S,2E)-1-isopropyl-4-((2S)-2-{(1R,2R)-1-methoxy-2-methyl-3-[(4R,5S)-4-methyl-2-oxo-5-phenyl-1,3-oxazolidin-3-yl]-3-oxopropyl}pyrrolidin-1-yl)-3-methyl-4-oxobut-2-enyl]-N¹,3-dimethyl-L-valinamideor a pharmaceutically acceptable salt thereof.
 45. A pharmaceuticalcomposition comprising an effective amount of a compound as claimed inclaim 1 in combination with one or pharmaceutically acceptable carriers.